Technical Library

Recent Entries in Technical Library

• Will the NHEM.f (C-12400) and NHEM.f M2 (C-12402) also grow in PromoCell Melanocyte Growth Medium M3?

  Yes, NHEM.f and NHEM.f M2 also grow in our optimized Melanocyte Growth Medium M3 (C24310) and can achieve > 15 population doublings.

  Related Links and Documents

  • Melanocyte Cell Culture
• I have a question regarding your Alanine Aminotransferase Activity Kit (PK-CA577-K752). What is the activity of the ALT Positive Control?

  The activity of the positive control is not determined. However, we can give you the OD value for the positive control from a specific lot. Please feel free to contact and share with us the lot number if you would like to see the expected OD value.

  Related Links and Documents

  • Alanine Aminotransferase (ALT/SGPT) Activity Assay Kit
• Following your protocol in a 96-well plate, how thick is the resulting Alginate Hydrogel matrix usually? And what are the maximum dimensions of an alginate hydrogel which still allow a sufficient exchange of nutrients and oxygen?

  50 µl of Alginate Hydrogel 3D Cell Culture Matrix per well (96-well plate format) is what is recommended. We have not measured to see what the exact thickness of this is. But this is enough to provide sufficient exchange of nutrients and oxygen. Thicker matrices may not be advisable but you may try.

   

  Related Links and Documents

  • Alginate Hydrogel 3D Cell Culture Matrix
• Does the BME 3D Cell Culture Matrix (PK-CA577-K518-5) contain laminin-1?

  The BME (Basement Membrane Matrix; animal-based) is rich in extracellular matrix proteins, including laminin (a major component), Collagen IV, heparin sulfate proteoglycans and a number of growth factors.

   

  Related Links and Documents

  • BME 3D Cell Culture Matrix
• According to literature, the NOS seems to need 3 or 5 cofactors to function, but the Nitric Oxide Synthase Activity Assay Kit contains only two, i.e. NOS Cofactor 1 and 2.

  The Cofactor Solutions are not a single reagent solution, but a mix of different cofactors needed for NOS reaction to proceed. Unfortunately, further information is proprietary and therefore cannot be disclosed.

  Related Links and Documents

  • Nitric Oxide Synthase (NOS) Activity Assay Kit I
• I have a question about the human fibronectin included in your Cell Invasion Assay Kit (PK-CA577-K925): What is the origin of the FN?

  The fibronectin is derived from human plasma and is produced in an animal-component free process.

  Related Links and Documents

  • Cell Invasion Assay Kits
• How much whole blood is required for 3 ml of the Lymphocyte Separation Medium in a 15 ml conical tube?

  In general the 15 ml tubes are filled with 3 ml Lymphocyte Separation Medium 1077 and the 50 ml conical tubes with 15 ml Lymphocyte Separation Medium.

  For the 15 ml tubes, it is recommended to use 3-8 ml, for the 50 ml tubes 15 – 30 ml of diluted* whole blood.

  *1:2 -1:3 in PBS

  Related Links and Documents

  • Lymphocyte Separation Medium 1077
• How do I get maximum expansion of the undifferentiated CD34+ cells? Is there a maximum cell density?

  The following is what we do here in our research lab for maximum expansion of the undifferentiated cells:
  – Seed the vial of cells at a density of 5,000 cells/ml as per PromoCell protocol (see Manual of human CD34⁺ progenitor cells).
  – Use our Hematopoietic Progenitor Cell Expansion Medium XF (C-28021) supplemented with our Cytokine Mix E.
  – Allow the cells to grow for one week. It may take 4-5 days for the cells to noticeably proliferate, but they will grow rapidly after this.
  – After the first week, change 2/3 of the media and culture for a second week.
  You should see up to a 200-fold expansion of the cells within this time. Cell density is not the major concern when expanding the progenitors. The media used and time in culture are more important factors. Some researchers expand the progenitors out a third week but we don’t recommend this for the health of the cells.

  Related Links and Documents

  • Hematopoietic Stem Cell Culture
• I have two questions regarding the PCR Mycoplasma Test Kit I/C. It is recommended to use a 1.5% agarose gel for the analysis. Would it also be possible to use a 2% or 3% pre-cast agarose gel? And do you recommend to use a DNA ladder?
  1. It is recommended to use a 1.5% agarose gel for the analysis of the amplified DNA. Theoretically, it is also possible to use a 2% or 3% pre-cast agarose gel as long as it is running for a sufficient distance to separate the bands at ∼270 bp (positive control and mycoplasma- positive samples) and 479 bp (internal control).
  2. A DNA ladder is not mandatory but is recommended to correctly identify all bands. We recommend using a 100 bp ladder, but a 250 bp ladder or similar, which allows you to identify the 270 and 479 bp bands is also suitable.

  Related Links and Documents

  • PCR Mycoplasma Test Kit I/C
• Which human AB serum does PromoCell recommend to generate M0 macrophages?

  We recommend to use Human Serum AB „off-the-clot”.

  Related Links and Documents

  • Macrophage Base Medium DXF
  • Application Note: Convenient large-scale differentiation
• I would like to differentiate hMSC into bone cells and further maintain the cells in culture after differentiation. Which medium should I use?

  Osteogenic differentiation of hMSCs with PromoCell MSC Osteogenic Differentiation Medium takes ∼12-14 days (please view the respective Application Note for a detailed protocol). Use fibronectin-coated plates and change the medium every third day.

  The bone cells tend to detach from the plastic after approximately 2 weeks, when differentiation is complete. Therefore, the tests should be performed promptly and the cells should be maintained in MSC Osteogenic Differentiation Medium until then.

  Related Links and Documents

  • Application Note: Osteogenic Differentiation and Analysis of MSC
  • human Mesenchymal Stem Cells
  • Mesenchymal Stem Cell Osteogenic Differentiation Medium
• I have purchased ACE2 blocking peptide and ACE2 antibody. Can you please tell me at what concentration to use the blocking peptide to block the antibody?

  Please find attached our general Western Blotting protocol as a guide.

   

   

  Related Links and Documents

  • General Western Blotting protocol
• I am performing MSC chondrogenic, adipogenic, and osteogenic differentiation and am staining the cells. The protocol says to use Saccomanno Fixation Solution. Is there an alternative method to fix the cells because I don’t have Saccomanno Fixation Solution?

  Yes, you can use 4.5% neutral buffered formalin. Paraformaldehyde should work as well.

   

  Related Links and Documents

  • Mesenchymal Stem Cell Culture
• Is PromoCell’s Cryo-SFM produced under GMP standard?

  No, our Freezing Medium Cryo-SFM (C-29910) is not produced under GMP Standard. It is for in vitro research use only and not appoved for diagnostic or therapeutic procedures.

  Related Links and Documents

  • Freezing Medium Cryo-SFM
• Is it possible to differentiate M1 macrophages from PBMC in 96-well plates?

  At PromoCell, we have not tested macrophage differentiation in 96-well plates, but we know from users that it is possible.

  The mononuclear cells also differentiate very well in the 96-well format. A plating density of 1 million PBMCs (without prior determination of monocyte content) per well has been shown to be optimal. The working volume in a 96-well plate is usually 100 µl.

  Related Links and Documents

  • M1-Macrophage Generation Medium DXF
• Can the medium for M2 macrophages be switched into RPMI or RPMI + M-CSF after differentiation?

  Unfortunately, we did not test this in our hands, and it must be tested by the customer. In fact, our medium is completely different from RPMI and therefore we cannot predict if this is working. We only know the successful long-term culture from our system with our media.

  Related Links and Documents

  • M2-Macrophage Generation Medium DXF
• Why does the macrophage differentiation in the PromoCell protocol take 10 days? In the literature or following the feedback from other researchers, it usually only takes 7 days.

  M1 / M2 polarization also takes seven days in the PromoCell system but the protocol contains two more days for optional macrophage activation. If you only want non-activated M1 / M2 macrophages, the process is usually completed after 7 days.
  Nevertheless, PromoCell does not recommend shortening the 10-day protocol because you actually get a plus in viability and cell yield (due to the re-attachment of floating cells) on day 8-10 due to the media change.

  Related Links and Documents

  • Application Note: Differentiation of M1- or M2-Macrophages
  • M1-Macrophage Generation Medium DXF
  • M2-Macrophage Generation Medium DXF
• I differentiated M1 macrophages from self-sourced PBMCs following your differentiating protocol. In the final cell culture, I can see small cells attached to the macrophages which are CD3+ (T-cells). What is the reason for the T-cell contamination after the differentiation?

  The reason for the higher number of lymphocytes in the macrophage culture is probably due to an insufficient washing step during the purification of the monocyte via adherence. The 3 washing steps in our protocol are essential to receive a monocyte population of over 90%.

  Related Links and Documents

  • Application Note
  • M1-Macrophage Generation Medium DXF
• How long do I have to activate the cryopreserved macrophages to measure cytokine release?

  In general, we recommend activating the cells for 24 hours or at least over night for all kind of activations. If the activation is not optimal in your experimental setting, you can increase or decrease the activation time accordingly.

   

  Related Links and Documents

  • Application Note
  • Macrophage Cell Culture
• I would like to analyze the macrophages by microscope and I would therefore have to plate the cells on fibronectin-coated glass. Did you test if the macrophages grow on fibronectin-coated glass?

  We did not test if the macrophages attach on fibronectin-coated glass.

  Related Links and Documents

  • Macrophage Cell Culture
• Following the recommended seeding density for your M1 macrophages (100,000 cells per cm2), I would need too many cells to have a complete 96- or 384-well plate. This is too expensive for me, can I reduce the number of cells per cm2?

  The recommended seeding density with 100,000 cells per cm² is needed for a confluent cell layer as the cells do not proliferate.
  However, you can reduce the seeding density by the factor 3 to 5 and the macrophages are still viable.

  Related Links and Documents

  • Macrophage Cell Culture
• Do you recommend a certain negative control for the activated macrophages to compare the cytokine release with?

  Even non-activated macrophages do release a certain amount of cytokines. Furthermore, you would have to be sure that the release of a certain cytokine is a direct consequence of the activation. Therefore, we do not think it is possible to have a general negative control for the cytokine release.

  Related Links and Documents

  • Macrophage Cell Culture
• Does PromoCell have any data regarding the cytokine profile and different markers of the M1/M2 macrophages after activation?

  No, we do not provide data about the cytokine profile of our M1/M2 macrophages after activation and we do not provide any further data beyond the scope of our quality control.

  Related Links and Documents

  • Macrophage Cell Culture
• Which cytokine concentrations does PromoCell recommend for activating M1/M2 macrophages?

  You may find all information regarding the activation and the cytokine concentrations in table 1 (page 5) of our Application Note.

  Related Links and Documents

  • Application Note
  • Macrophage Cell Culture
• Did PromoCell try plating the cryo-macrophages in 96-well plates?

  We did not plate the cryo-macrophages in 96-well plates. However, we heard from other customers that they have successfully used our macrophages in this kind of plate.

  Related Links and Documents

  • Macrophage Cell Culture
• Is the Aromatase Assay Buffer compatible with a classical BCA protein quantification kit (e.g. Pierce™ BCA Protein Assay Kit)?

  The assay buffer of our Aromatase Activity Assay Kit (PK-CA577-K983) has 50 mM potassium phosphate and may not be compatible with BCA. But it is compatible with Bradford assay.

  Related Links and Documents

  • Aromatase (CYP19A) Activity Assay Kit
• What MATra reagent does PromoCell recommend for the transfection of vascular Smooth Muscle Cells?

  Bovine carotid artery smooth muscle cells have been successfully transfected using MATra-A.

  Related Links and Documents

  • Magnet-Assisted Transfection
• What’s the difference between PromoFectin (PK-CT-2000-100) and PromoFectin-siRNA (PK-CT-2000-RNA-200)?

  Both, PromoFectin and PromoFectin-siRNA, are non-liposomal transfection reagents.

  • Our (broad-range) PromoFectin can be used to transfect primary cells and cell lines with DNA or RNA.
  • PromoFectin-siRNA was developed to specifically deliver siRNA into a wide variety of cell types.

  Related Links and Documents

  • PromoFectin reagents
• Can I spray the Spore-EX Spray on plexiglass surfaces without damaging it?

  This spray is compatible with plastic/plexiglass. However it will leave residue as it is not an alcohol based formulation, so you may need to wipe dry with a paper towel.

  Related Links and Documents

  • Spore-EX Spray
• I received 3 DetachKits yesterday. Some of the tubes have different colors, although they all come from the same batch.

  The components of the PromoCell DetachKit may occasionally arrive with a non-uniform color appearance. This phenomenon is known to PromoCell’s Quality Assurance Dept.
  It is reversible and has no influence on the quality of the product.

  Related Links and Documents

  • PromoCello DetachKit
• Are endothelial cells α-SMA-negative under all circumstances?

  Scientific findings from different groups, as well as our own results indicate that the presence or absence of α-SMA is not a valid indicator for the composition of a given cell population. Research findings and our inhouse data suggest that endothelial cells are not α-SMA-negative under all circumstances. α-SMA negativity is not an intrinsic property of endothelial cells but can vary depending on extrinsic influences.

  Lot-specific values for α-SMA in our Certificates of Analysis (CoAs) are therefore not considered meaningful.

  Related Links and Documents

  • Endothelial Cell Culture
  • Ando et al.; Am J Physio l. 1999 May;276(5):H1755-68
  • Lu et al.; Stem Cells Dev . 2004 Oct;13(5):521-7
  • Azuma et al.; Biochem Biophys Res Commun . 2009 Mar 13;380(3):620-6
  • Frid et al.; Circ Res . 2002 Jun 14;90(11):1189-96
  • Kovacic et al.; J Am Coll Cardiol . 2019 Jan 22;73(2):190-209
  • Jones et al.; Am J Respir Cell Mol Biol . 1999 Apr;20(4):582-94
  • Ma et al.; Front Cell Dev Biol . 2020 Apr 21;8:260
  • Kocher and Madri; In Vitro Cell Dev Biol . 1989 May;25(5):424-34
  • Amberger et al.; FEBS Lett . 1991 Aug 5;287(1-2):223-5
• Is vWF a great marker for Endothelial Cells?

  Secretory granules called Weibel-Palade bodies (WPBs) containing Von Willebrand factor (vWF) are more linked to the formation of a confluent endothelial monolayer. Research shows that vWF expression is dynamic and highly dependent on the cell culture conditions such as confluence and passage number.
  Therefore, having vWF as a quality control marker for each lot of ECs is not really necessary.

  Related Links and Documents

  • Endothelial Cell Culture
  • Howell et al.; Mol Membr Biol. Nov-Dec 2004;21(6):413-21
• The components of my DetachKit show different colors. Is this normal or does it affect their quality?

  The components of the PromoCell DetachKit may arrive on occasion with a non-uniform color appearance. This phenomenon is known by PromoCell’s Quality Assurance. It is reversible and does not influence the quality of the product.

  Related Links and Documents

  • PromoCell DetachKit
  • PromoCell DetachKit 2
• The supplements arrived defrosted. Are they still ok?

  PromoCell supplements (SupplementMixes and SupplementPacks) are routinely shipped with dry ice. On occasion, the supplements may arrive defrosted. You can either use them immediately to prepare fresh growth media or refreeze them at -20°C. We have performed extensive stability tests and can guarantee that the supplements are stable for up to 1 week at room temperature.

  Sole exception: The SupplementMix (C-39420) of our Melanocyte Growth Medium M2 is less stable and should no longer be used when it arrived thawed, at room temperature.

  Related Links and Documents

  • Media for Human Primary Cell Culture
  • Melanocyte Growth Medium M2
• Can you please let me know whether Spore-EX Spray can eliminate DNA and RNA from surfaces?

  Yes, our Spore-EX Spray (PK-CC91-5053) is also suitable for denaturing DNA and RNA.

  Related Links and Documents

  • Spore-EX Spray
• What is the Endotoxin level of PromoCell’s PBS?

  The specification of endotoxin level is < 1 EU/ml.
  For all our produced batches, the result is 0.005 EU/ml.

  Related Links and Documents

  • Dulbecco’s PBS
• Can I defrost Accutase Solution, prepare aliquots and refreeze them?

  Yes, Accutase Solution can be defrosted, aliquoted, and then refrozen.

  Defrosting: Accutase should be defrosted overnight in the refrigerator or placed in a tub of cold tap water.  Do not defrost in a 37°C water bath.
  Stability: Once thawed, it is stable for at least 2 months in the refrigerator if stored promptly after use.

  Related Links and Documents

  • Accutase Solution
• What is the difference between the PCR Mycoplasma Test Kit I/C (PK-CA91-1024) and PCR Mycoplasma Test Kit II (PK-CA20-700-20)?

  Both PCR Mycoplasma Test Kits are designed for conventional PCR but differ in the following points:

  • Format: The reagents are delivered lyophilized in Test Kit I/C, liquid (5x master mix) in Test Kit II.
  • Detection Spectrum: The spectrum of detected strains is slightly different: Test Kit II also detects e.g. M. pneumoniae and Spiroplasma citrii (which are both mentioned in the EUR. PHARM. as test/reference species) and a few others, which are however of no importance in cell cultures.
  • Size of internal controls:
    – PCR Mycoplasma Test Kit I/C ⇒ internal control: 479 bp, positive control: 270 bp
    – PCR Mycoplasma Test Kit II ⇒ internal control: 357 bp, positive control: 270 bp

  Related Links and Documents

  • PCR Mycoplasma Test Kit I/C
  • PCR Mycoplasma Test Kit II
• Is it possible to refreeze the hCD34+ progenitor cells after having amplifed them in PromoCell HPC Expansion Medium?

  Yes, it is possible to refreeze them.

  Related Links and Documents

  • CD34+ Progenitor Cells
  • HPC Expansion Medium DXF
• Is it possible to refreeze PromoCell’s Macrophages?

  Once thawed, our assay-ready human M1- and M2 Macrophages cannot be refrozen.

  Related Links and Documents

  • Human M1 Macrophages (GM-CSF) monocyte-derived
  • Human M2 Macrophages (M-CSF) monocyte-derived
• Why are the cryopreserved macrophages abbreviated as hMDM-GMCSF(-) and hMDM-MCSF(-), respectively? What does the (-) stand for?
  • hMDM-GMCSF is the abbreviation for human monocyte-derived macrophages. They are polarized [⇒ differentiated with GM-CSF] but non-activated [⇒ (-)] M1 macrophages. 
  • hMDM-MCSF is the abbreviation for human monocyte-derived polarized [⇒ differentiated with M-CSF] but non-activated [⇒ (-)] M2 macrophages. 

  The macrophages can be seeded into all kinds of TC vessels. After plating, they can be maintained as biologically functional adherent cultures for several weeks.Optionally, user-customizable activation of the cells can be performed. For details, please see attached Application Note, page 2, Fig. 3 and page 5, Tab. 1).

  Related Links and Documents

  • Application Note: Assay-ready adherent cultures of fully functional human primary macrophages using cryopreserved cells
  • Cryopreserved Macrophages
  • Macrophage Generation Media DXF
• What do you recommend to use to loosen the monocyte fraction from the tissue culture plate (after attachment with the Monocyte Attachment Medium) in order to assess phenotypic/functional markers by flow cytometry?

  Our Macrophage Detachment Solution (C-41330) can be used to detach monocytes from the tissue culture plastic.

  Related Links and Documents

  • Monocyte Attachment Medium
  • Macrophage Detachment Solution
• If I isolate fresh CD14-monocytes for M1/M2 macrophage generation, can I culture them for a period of time before differentiation?

  This is not advised. Please seed the freshly isolated CD14-monocytes immediately in the Monocyte Attachment Medium. Adding a culturing step will change the biological characteristics of monocytes very rapidly.

  Related Links and Documents

  • Application Note: Differentiation of M1- or M2-Macrophages from PBMC/Monocytes
  • Monocyte Attachment Medium
• Can you provide a list of cell lines that have been successfully used for tumorsphere formation with the PromoCell 3D Tumorsphere Medium XF?

  The following cell lines have been tested at PromoCell to form tumorspheres in 3D Tumorsphere Medium XF (C-28070):

  • U-87 MG
  • MCF-7
  • MDA-MB-231
  • HT-29
  • HT1080
  • HepG2
  • A-549
  • Panc-1
  • LNCaP
  • A-431

  ⇒ For more details, please view the attached Application Note.
  In addition, we have received customer feedbacks for the following cell lines:

  • HCT-116 (human colorectal carcinoma cell line)
  • Capan-1 (human pancreatic adenocarcinoma cell line)
  • PC3 (human prostate cancer cell line)
  • C42B (osteotropic prostate cancer cell line)
  • NCI-H23 (human lung epithelial adenocarcinoma cells)
  • IMR-32 (human neuroblast cell line)
  • A818-6 (human pancreatic ductal adenocarcinoma cell line)
  • HEK293 (human embryonic kidney cells)
  • Calu-1 (non-small-cell lung cancer cell line)

  Related Links and Documents

  • Application Note: Tumorsphere Culture of Cancer Stem Cells (CSC) with the PromoCell 3D Tumorsphere Medium XF
  • 3D Tumorsphere Medium XF
• I have a question regarding the protocol of tumorsphere culture using PromoCell’s 3D Tumorsphere Medium XF (see Application Note, p. 5, step 5.B): What will happen if the spheroids are collected using centrifugation instead of gravity sedimentation?

  To collect the tumorspheres prior to passage, a centrifugation step at 350xg can be applied instead of gravity sedimentation.
  When using centrifugation, please be careful when aspirating the supernatant because the pellet can be loose. Moreover, centrifugation can lead to subsequent dissociation of the spheres. 
  The advantage of the sedimentation method is that – together with the supernatant – debris will be removed from the spheres which results in “clean” tumorsphere cultures.

  Related Links and Documents

  • 3D Tumorsphere Medium XF
• Is trypsin, accutase, or any other enzymatic detachment reagent used during the production of PromoCell’s cryopreserved macrophages?

  The production of PromoCell’s cryopreserved macrophages is performed under consistently defined culture conditions and excludes the use of any enzymatic and/or non-defined detachment reagents.

  Related Links and Documents

  • Application Note: Assay-ready adherent cultures of fully functional human primary macrophages using cryopreserved cells
  • Macrophages
• Can PromoCell MSC Growth Medium 2 (C-28009) be used for Mesenchymal Stem Cells from rat?

  Yes, we have received a customer feedback that our MSC Growth Medium 2 also works for rat MSCs.
  The rat cells grow nicely in this medium and have a good viability.

  Related Links and Documents

  • MSC Growth Medium 2
• Which medium does PromoCell recommend for endothelial cell transfection ?

  There are several factors that can influence successful transfections, e.g. viability and density of the cells, choice of the transfection reagent, quality and type of the transfected molecules (plasmids, siRNA, oligonucleotides), as well as the culture medium and supplements used.
  When using PromoCell Endothelial Cell Growth Media for cell transfection, please follow the instructions below:

  Heparin, which is included in our Endothelial Cell Growth Medium (C-22010/C-22110), Medium 2 (C-22011/C-22111) and Medium MV (C-22020/C-22120) may reduce the transfection efficiency.  We therefore recommend to use our heparin-free Endothelial Cell Growth Medium MV2 (C-22022/C-22121). Alternatively, you may use Endothelial Cell Growth Medium Kit (C-22110), Medium 2 Kit (C-22111), or Medium MV Kit (C-22120) without adding the ECGS/heparin supplement to the Basal Medium.

  Please note: Before and after transfection, the cells should be cultured in complete Growth Medium including heparin to ensure optimal growth.

  Related Links and Documents

  • Manual: Endothelial Cell Growth Medium/Medium 2
  • Manual: Endothelial Cell Growth Medium MV/ MV2
  • Cell Transfection reagents
• Are there PromoCell specialized media / primary human cells alternatives to other supplier’s products?

  Please see our attached alternative product guides for media and cells.

  Please note: Our media do not contain antibiotics. For optimal cell growth, we recommend to refrain from using antibiotics. However, when a sterile environment cannot be 100% ensured, it may be advisable to add antibiotics to your media so as to protect the cultures from potential microbial infections.

  Related Links and Documents

  • Alternative Product Guide - Primary Human Cells: Lonza® – PromoCell
  • Alternative Product Guide - Primary Human Cell Media: Lonza® – PromoCell
  • PromoCell's packaging systems: Medium (Ready-to-use) vs. Medium Kit
• Do I have to use coated tissue culture vessels for culturing PromoCell Normal Human Cells?

  It is not necessary to use coated flasks for (most of) our Normal Human Cells but it can be done. As coating with extracellular matrix proteins can affect cellular metabolism, it is recommended to use the same coating material for a complete set of experiments.
  Cells that need to be grown on coated dishes:

  • Mesenchymal Stem Cells (C-12974/C-12971/C-12977) need Fibronectin-coating when grown in PromoCell MSC Growth Medium XF (C-28019) and when differentiated in MSC Neurogenic (C-28015), Adipogenic (C-28016), or Osteogenic (C-28013) Differentiation Media.
  • Human monocyte-derived macrophages (C-12914/C-12916/C-12915/C-12917) must be seeded into Fibronectin-coated culture vessels in combination with PromoCell’s M1- and M2-Generation Media XF (C-28055, C-28056).
  • For efficient induction of osteoblast mineralization with PromoCell’s Osteoblast Mineralization Medium (C-27020), the TC plates should be pre-coated with collagen type I.

  Related Links and Documents

  • Mesenchymal Stem Cell Culture
  • Macrophage Cell Culture
  • Osteoblast Cell Culture
  • Fibronection Solution
• Why is it not necessary to use Monocyte Attachment Medium (C-28051) when using DC Generation Medium XF (Ready-to-use, C-28052)?

  The DC Generation Medium XF (C-28052) is recommended for freshly isolated monocytes or mononuclear cells. When using this medium, cells will attach efficiently, so that an additional medium like the Monocyte Attachment Medium (C-28051) is not necessary.
  In contrast, when using the DC Generation Medium (C-28050) for freshly isolated monocytes or mononuclear cells, the Monocyte Attachment Medium (C-28051) is needed for an efficient attachment.

  Related Links and Documents

  • Application Note: Generation of monocyte-derived Dendritic Cells (moDCs)
  • Monocyte Attachment Medium
  • Dendritic Cell Generation Medium
  • Dendritic Cell Generation Medium XF
• Does the PromoCell 3D Tumorsphere Medium XF (C-28070) maintain the stem cell character of cancer stem cells?

  During our quality control we do not determine the presence or maintenance of respective stem cell markers in the tumorspheres. We use a functional approach instead, by culturing the tumorspheres over serial passages in C-28070. Tumorsphere formation requires the biological features of Anoikis resistance and self-renewal and therefore indicates the presence of cancer stem cells (CSC)/cancer initiating cells (CIC) in the spheres. 3D tumorsphere culture thus allows scientists to study the biology of CSCs without any background knowledge on CSC markers.

  Related Links and Documents

  • Product Manual: 3D Tumorsphere Medium XF
  • Application Note: Tumorsphere Culture of Cancer Stem Cells (CSC) with the PromoCell 3D Tumorsphere Medium XF
  • Application Note: Determination of the Tumorsphere Formation Efficiency (TFE) with the PromoCell 3D Tumorsphere Medium XF
• What are the characteristic markers of HDLEC (C-12216, C-12217) and HDBEC (C-12211, C-12225)?
  • HDLEC and HDBEC both express the typical endothelial cell marker CD31 (= PECAM-1). 
  • HDLEC cultures are additionally tested positive for podoplanin, a transmembrane glycoprotein involved in lymphatic vessel formation, whereas HDBECs are podoplanin-negative.

  Related Links and Documents

  • Microvascular Endothelial Cells
• Can I use the 3D Tumorsphere Medium XF for mouse cell lines?

  Yes, we have received a customer feedback that our 3D Tumorsphere Medium XF (C-28070) has been successfully used for tumorsphere formation of mouse cell lines.

  Related Links and Documents

  • Tumorsphere Culture of Cancer Stem Cells (CSC) with the PromoCell 3D Tumorsphere Medium XF
  • 3D Tumorsphere Medium XF
• What’s the chemical basis of Mycoplasma-ExS Spray? How does it work?

  Mycoplasma-ExS Spray uses a silver-based technology to eliminate microbes. It contains active polymerically stabilized ionical silver chloride (AgCl) that is UV-stable. 
  The silver chloride forms a thin film on any surface, providing an immediate antimicrobial effect. Moreover, it can be also used for long-term elimination because the silver and chloride ions are released extremely slow from the silver polymer layer.
  Mycoplasma-ExS is effective against a broad spectrum of unwanted microbes commonly found in biological labs including Mycoplasma, Pseudomonas aeruginosa, Escherichia coli, Enteroccocus hirae, and Staphylococcus aureus. 

  Related Links and Documents

  • Mycoplasma-ExS Spray
• When should I use phenol red-free basal media for cell culture?

  It has been shown that phenol red has estrogenic properties. Phenol red-free media are therefore generally used in studies evaluating steroid hormone action in cultured, estrogen-responsive cells (Berthois et al., 1986). Furthermore, phenol red can also interfere with some analytical methods like photometric analyses.

  Related Links and Documents

  • Berthois et al.; Proc Natl Acad Sci U S A. 1986 Apr;83(8):2496-500
• Where can I find the composition of your basal media?

  The formulation of our basal media is proprietary information. If you need to know the concentration of a particular component for your experiments, please contact the PromoCell Technical Customer Service.

• Where can I find the composition of the supplements?

  The qualitative and quantitative composition of the supplements can be found on our website and in the data sheets of the specialized media. When there is no such information specified, the composition of the supplements is confidential.

• What antibiotics concentration should I add to the PromoCell media?

  If antibiotics are deemed necessary the recommended final concentrations are:
  100 U/ml penicillin + 100 µg/ml streptomycin or
  50 µg/ml gentamicin + 50 ng/ml amphotericin B

  Please note: Addition of antibiotics can reduce the growth rate of the cells.

• Are the Renal Epithelial Cells isolated from proximal or distal tubuli?

  PromoCell provides two types of Renal Epithelial Cells: Human Renal Epithelial Cells (HREpC) and Human Renal Cortical Epithelial Cells (HRCEpC).
  HREpC are isolated from the adult kidney and stain positive for cytokeratin. They comprise a heterogeneous population of renal epithelial cells. HRCEpC are isolated from the cortex of the kidney and comprise cells from proximal and distal tubuli. They also stain positive for cytokeratin.

  Related Links and Documents

  • Epithelial Cells
• How does PromoCell characterize their kidney cells (HREpC; HRCEpC) ?

  Our kidney cells are characterized by their epithelial cell morphology and by cytokeratin expression using a pan-cytokeratin antibody.

  Related Links and Documents

  • Renal epithelial cells
• What is the difference between PromoCell’s HREpC and HRCEpC?

  Our HREpC (C-12665) comprise a heterogenous mixture of renal epithelial cells isolated from cortex and medulla. The HRCEpC (C-12660) are derived from renal cortex only.

  Related Links and Documents

  • Renal Epithelial Cells
  • Schematic structure of a kidney
• How many cells should I use for one Mycoplasma EX treatment to sucessfully remove Mycoplasma from my culture? Do you recommend a specific cell number for the treatment?

  According to our protocol 5 ml cell suspension is added to 4.5 ml of Mycoplasma-EX reagent dissolved in growth medium (e.g. in a T25 flask).  
  The recommended cell number depends on the one hand on the specific cell type and the size of the culture flask you use. Mycoplasma is often located on the cell surface of the cells. The protocol (page 4 of the Instruction Manual) says to split the cells as thin as possible so that the number of transferred mycoplasma is reduced before the initial treatment.
  If e.g. you normally use split ratios between 1:6 and 1:10 for your particular cell type, then rather use 1:10 before starting the Mycoplasma EX treatment. If using seeding densities between 5.000 and 10.000 cells/cm², then rather use 5.000 cells/cm².

  Related Links and Documents

  • Instruction Manual: Mycoplasma-EX Kit
  • Mycoplasma-EX
• What could be the reason for a lower yield when using M2 Macrophage Generation Medium compared to M1 Macrophage Generation Medium for a culture started with the same monocytes?

  For the M2 Macrophage Generation Medium, it is extremely important that the shelf life of 2 weeks (after addition of the cytokines) is not exceeded; the yield will quickly decrease thereafter. It is best to use the M2 medium as fresh as possible to avoid discrepancies between M1 and M2 yield.

  Related Links and Documents

  • Application Note: Differentiation of M1- or M2-Macrophages from PBMC/Monocytes
  • Macrophage Generation Media DXF
• Which products can I use to clean laminar flows and to prevent contamination, especially Mycoplasma?

  We recommend to disinfect the laminar flow 1-2x per day with one of the following disinfection sprays:

  • Mycoplasma-ExS Spray: effective against bacteria (especially Mycoplasma) and fungi within 5 min ⇒ basic level disinfectant
  • Spore-Ex Spray: effective against bacteria (including Mycoplasma), fungi, viruses and spores within 1 min ⇒ highest level disinfectant

  Related Links and Documents

  • Mycoplasma-ExS Spray
  • Spore-EX Spray & Wipes
• Can the PromoKine 3D Cell Culture Harvesting Kit also be used to isolate cells from Collagen or Laminin matrices?

  Yes, it is possible. The kit is matrix agnostic. However, it might be necessary to optimize the incubation time.

  Related Links and Documents

  • Instruction Manual: 3D Cell Culture Harvesting Kit
  • 3D Cell Culture - Cell Harvesting
• It is mentioned in the Instruction Manual of the PCR Mycoplasma Test Kit I/C that the signal for the internal control DNA may fade with increased amount of mycoplasma contamination. Does this mean that the internal control is not visible anymore when the cells are heavily contaminated?

  The amplification of the internal control and the mycoplasma DNA is a competitive reaction: If the amount of Mycoplasma amplicons in the test sample is higher than 10⁴ copies/ml, then less internal control amplicons (fixed amount per test reaction tube) will be amplified, leading to a lower signal of the internal control band. In some cases, the internal control is actually not visible anymore.

  Interpretation of results:
  PCR setup	band at 479 bp	band at 270 bp	Interpretation
  negative control	+	—	internal control visible: correct PCR setup
  positive control	(+)	+	correct PCR setup; indicates size of mycoplasma band*
  sample	+	+	mycoplasma positive
  sample	(+)	++	strongly positive*
  sample	+	—	negative, no mycoplasma contamination
  sample	—	—	no internal control (479 bp) visible: PCR inhibition ⇒ use DNA extraction method**, re-test

  * Internal control band might not be visible in case of strong mycoplasma DNA amplification/mycoplasma contamination** Please see page 5/6 of Instruction Manual

  Related Links and Documents

  • Instruction Manual: PCR Mycoplasma Test Kit I/C
• What is the differentiation ratio that can be achieved with PromoCell’s subcutaneous HWP?

  Most of our subcutaneous preadipocyte lots achieve > 80-90% differentiation when differentiation is induced at P2 (directly after thawing).
  We generally recommend using cells for differentiation tests that haven’t undergone more than 4-5 doublings (a maximum of 1 passage after thawing), as the differentiation ratio will decline with the age of the cells.

  Related Links and Documents

  • Human White Preadipocytes
  • Preadipocyte Growth Medium
  • Preadipocyte Differentiation Medium
  • Adipocyte Nutrition Medium
• What’s the principle of Mycoplasma Test Kit I/C? Does the Kit include Taq Polymerase?

  Mycoplasma Test Kit I/C is based on the amplification of the highly conserved 16S rRNA region of the Mycoplasma genome. The PCR components (including Hot-Start Taq Polymerase) come lyophilized in 0.2 ml PCR tubes. For setting up the PCR, simply cut off the number of tubes required, rehydrate with rehydration buffer and add the sample. The Kit includes internal control DNA (as a negative control and for verifying a successful PCR run) as well as a positive control.
  A successfully performed PCR reaction is indicated by a 479 bp band (internal control) on the agarose gel. The positive control yields a band at 270 bp. PCR Mycoplasma Test Kit I is also available as Kit for Real Time PCR (PK-CA-91-3025C, PK-CA-91-3050C).

  Related Links and Documents

  • PCR Mycoplasma Test Kit I/C
  • PCR Mycoplasma Test Kit I/RT
• Can you provide some information about the internal control of the PCR Mycoplasma Test Kit I/C (source, sequence etc.)? Is it a mixture or a specific DNA?

  The internal control consists of a non-coding synthetic DNA. The exact sequence is confidental. It is a specific DNA, not a mixture and shows one distinct band of 479 bp in the agarose gel after PCR amplification.

  Related Links and Documents

  • PCR Mycoplasma Test Kit I/C (for conventional PCR)
• After detaching the macrophages for flow cytometric analysis, I noticed that the washing and centrifugation steps take a long time. Can I shorten the spin time to 10 minutes at 350 x g?

  Spinning the cells for 15 min at 350 x g has been proven and tested by PromoCell Research & Development. The QC department uses these settings during testing. Any lower centrifugation value (g-force and/or time) will lead to significant cell loss by means of non-sedimented, but intact, macrophages.

  Related Links and Documents

  • Differentiation of M1- or M2-Macrophages from PBMC/Monocytes
  • Macrophage Generation Media DXF
  • Macrophage Detachment Solution DXF
• What is the difference between the MATra-A Reagent and the MATra-S Immobilizer?

  1. The MATra-A reagent is for adherent cells: The nucleic acid only needs to be combined to MATra-A and magnet-assisted transfection can be performed.
  2. MATra-S Immobilizer is needed for suspension cells: The cells are made adherent by incubating them with the magnetic reagent MATra-S Immobilizer. Then MATra-A loaded with the nucleic acid is applied to the culture and magnet-assisted transfection can be performed.

  Related Links and Documents

  • MATra reagents
• When monitoring luciferase expression, can I add assay buffer to all my samples and read them simultaneously?

  No. PromoKine’ s Renilla and Firefly Assay Kits (PK-CA707-30003; PK-CA707-30004; PK-CA707-30081) are based on flash luminescence. Samples should be treated and read individually. Alternatively, you may purchase the Firefly Luciferase Reporter HTS Kit (PK-CA707-30028). This Assay Kit is a proprietary mixture of substances that modify the enzymatic reaction to produce a long lasting signal (steady glow) by preventing the formation of adenyl-oxyluciferin at the enzyme surface.

  Related Links and Documents

  • Luciferase Reporter Assay Kit I (Firefly)
  • Luciferase Reporter Assay Kit II (Renilla)
  • Luciferase Reporter Assay Kit III (Firefly & Renilla Single-tube Assay)
  • Luciferase Reporter HTS Kit (Firefly)
• Why are my luminescence numbers different from those in the product information sheet?

  Luminescence values are relative and depend on the experimental system. For example, luminometers from two different manufacturers each may have 6-log capability, but one may measure a range of 1 to 1,000,000 relative light units (RLU) while the other measures 0.01 to 10,000 RLU. Both instruments can measure a dynamic range of 6-log units, but they read out different arbitrary units.

  Related Links and Documents

  • Luciferase Reporter Assay Kit I (Firefly)
  • Luciferase Reporter Assay Kit II (Renilla)
  • Luciferase Reporter Assay Kit III (Firefly & Renilla Single-tube Assay)
  • Luciferase Reporter HTS Kit (Firefly)
• Why would I need to use the Dual Luciferase Reporter Assay Kit (Firefly & Renilla Single-tube Assay)?

  Renilla is often used as an internal control for transfection efficiency while the Firefly luciferase vector is used for reporter activity. They also can be reversed, with Renilla as the reporter and Firefly as the internal control.
  PromoKine’s Luciferase Reporter Assay Kit III (PK-CA707-30081) allows measurement of Firefly and Renilla luciferase activity in the same sample with high sensitivity and linearity.

  Related Links and Documents

  • Luciferase Reporter Assay Kit III (Firefly & Renilla Single-tube Assay)
• Can I use Promega’s Lysis Buffer or Assay Buffer with PromoKine Luciferase Reporter Assay Kits?

  No. The buffers are based on different proprietary formulations.

  Related Links and Documents

  • Luciferase Reporter Assay Kit I (Firefly)
  • Luciferase Reporter Assay Kit II (Renilla)
  • Luciferase Reporter Assay Kit III (Firefly & Renilla)
  • Luciferase Reporter HTS Kit (Firefly)
• What’s the difference between Renilla and firefly luciferase?

  Firefly luciferase is a 62 kDa protein isolated from beetles (Photinus pyralis), while Renilla luciferase is a 36 kDa protein from sea pansy (Renilla reniformis).
  These enzymes differ in their substrate and cofactor requirements. Firefly luciferase uses luciferin in the presence of oxygen, ATP, and magnesium to produce light, while Renilla luciferase requires only coelenterazine and oxygen.
  Firefly luciferase produces a greenish yellow light in the 550–570 nm range. Renilla luciferase produces a blue light of 480 nm.
  These enzymes can be used in dual-reporter assays due to their differences in substrate requirements and light output.

  Related Links and Documents

  • Luciferase Reporter Assay Kits
• What is the half-life of the Renilla and Firefly Assays?

  PromoKine’ s Renilla and Firefly Assays (Luciferase Reporter Assay Kits I, II, and III) are flash luminescence assays. The approximate half-lives are 1-2 minutes for Renilla and 10 minutes for Firefly. The Luciferase Reporter HTS Kit has a half-life of about 3-5 hrs. However, the signal is about 1/10 of the flash luminescence assays.

  Related Links and Documents

  • Luciferase Reporter Assay Kit I (Firefly)
  • Luciferase Reporter Assay Kit II (Renilla)
  • Luciferase Reporter Assay Kit III (Firefly & Renilla)
  • Luciferase Reporter HTS Kit (Firefly)
• How can I measure the ratio of successful transfections?

  Transfection rates can directly be measured by (co-)transfecting a GFP-expressing plasmid. The percentage of cells that have taken up and expressed the foreign nucleic acid can be visualized with a fluorescence microscope (by green autofluorescence).
  Another way of quantification is to transfect luciferase or beta-galactosidase expressing reporter vectors and to determine the activity of these enzymes as a measure of successful transfection. Several commercial cell based reporter assays are available for this purpose (Firefly and Renilla Luciferase Reporter Assay Kits; GFP reporter Assay Kit; beta-Galactosidase Activity Assay Kit; all available from PromoKine).

  Related Links and Documents

  • Transfection Reagents
  • Reporter Assays
• What is the principle of the PromoKine Senescence Detection Assay?

  Cellular senescence limits the lifespan of mammalian cells, prevents unlimited cell proliferation and is thought to be a tumor suppressive mechanism. In the cell culture dish, senescence represents a state in which the cells remain viable, but loose the ability to divide. Furthermore, the cells display an increase in size, senescence-associated expression of beta-galactosidase (SA-beta-Gal) activity, and altered patterns of gene expression.

  • The PromoKine Senescence Detection Kit I is designed to histochemically detect SA-beta-Gal activity in fixed cultured cells and tissue sections. Senescent cells develop a blue color precipitate that can be observed under the microscope.
  • The Senescence Detection Kit II is designed to fluorescently detect SA-beta-Gal activity in cultured cells by FACS (FL1 channel)

  SA-beta-Gal is present only in senescent cells and is not found in presenescent, quiescent or immortal cells.

  Related Links and Documents

  • Senescence Detection Kit I
  • Senescence Detection Kit II
• Can the Senescence Detection Kit I be used with tissue sections?

  Yes, the kit has been used for skin sections successfully. Briefly, the tissue was frozen in liquid nitrogen, and mounted in OCT. The thin sections (4 µm) were cut, mounted onto glass slides, fixed in 1% formalin in PBS for 1 min at room temperature, washed in PBS, and immersed overnight in beta-Gal staining solution. Then you can view under bright field at 100-200x magnifications.
  The staining results can be found in the article below:Dimri et al.; PNAS 1995; 92(20):9363-7

  Related Links and Documents

  • Senescence Detection Kit I
  • Dimri et al. PNAS. 1995
• Could you send me a reference article describing SA-beta-Galactosidase as a senescence marker?

  The following paper describes beta-galactosidase as a biomarker for senescent human cells in culture:
  Dimri et al.; Proc Natl Acad Sci U S A. 1995 Sep 26;92(20):9363-7

  Related Links and Documents

  • Senescence Detection Kit I
  • Senescence Detection Kit II (fluorometric)
  • Dimri et al. PNAS. 1995
• Does the Senescence Detection Kit I detect transient expression of p53 (3-5 days) or longer term expression?

  The kit will detect senescent cells on the basis of senescence-associated expression of beta-galactosidase. It does not matter what causes senescence. If the p53 expressing cells become senescent, then the kit should detect them.

  Related Links and Documents

  • Senescence Detection Kit
• What is the principle of the Mitochondrial Apoptosis Staining Kit?

  Our Mitochondrial Apoptosis Staining Kit provides a rapid two-color staining procedure performed directly with live cells. The assay quantifies the changes in mitochondrial transmembrane potential, one of the earliest intracellular events upon induction of apoptosis. It uses a cationic fluorescent dye, MitoStain, that fluoresces differently in healthy versus apoptotic cells:

  • In non-apoptotic healthy cells, the dye exists as a monomer in the cytosol (green) but also accumulates and aggregates in the mitochondria, emitting a bright red fluorescence
  • In apoptotic and necrotic cells, due to disruption of the mitochondrial transmembrane potential, the fluorescent dye only exists as monomer and stains the cytoplasm green.

  Analysis of the stained cells can be done by FACS or fluorescence microscopy.

  Related Links and Documents

  • Mitochondrial Apoptosis Staining Kit
• Is it normal that CD34+ or CD133+ progenitor cells attach to tissue culture plastic after a week-long culture in HPC Expansion Medium XF (C-28021)?

  The cells sink down to the bottom of the culture vessel but don’t really attach. They retain a roundish morphology and can be rinsed off with culture medium easily.

  Related Links and Documents

  • Human CD34+ Progenitor Cells
  • Hematopoietic Progenitor Expansion Medium XF
• At what passage are PromoCell Human Blood Cells upon arrival?

  PromoCell Blood and Blood Progenitor Cells are cryopreserved directly after isolation (= P0). They haven’t been in culture before freezing.

  Related Links and Documents

  • hCD34+
  • hMoCD14+
  • hMNC
• Will the hematopoietic progenitor cells proliferate in Hematopoietic Progenitor Cell Expansion Medium XF (C-28021) without Cytokine Mix E (C-39890)?

  For the expansion of hematopoietic progenitors (CD133+ cells, CD34+ cells), PromoCell has developed Hematopoietic Progenitor Cell Expansion Medium XF (C-28021), a serum-free and xeno-free formulation. The medium must be supplemented with either Cytokine Mix E (C-39890) or with the user’s own mixture of cytokines. Cytokine Mix E is a ready-to-use mix containing recombinant human TPO, SCF, flt3-ligand, and IL-3.
  The strong expansion of the progenitor cells in Expansion Medium XF + Cytokine Mix E persists for at least two weeks, resulting in a 200-300x increase of the cell number with only 20-30% differentiated cells.
  Note: If starting with CD133+ cells, the CD133 marker is getting lost during this expansion step. The resulting cells are D34⁺/CD38^–/CD133^–.

  Related Links and Documents

  • hCD34+ Progenitor Cells
  • Hematopoietic Progenitor Expansion Medium XF
• How long does it take to differentiate CD34+ or CD133+ progenitor cells into hematopoietic cells when using PromoCell Hematopoietic Progenitor Medium (C28020)? How often should I change the medium during this process?

  The process of hematopoietic differentiation takes approximately 3 weeks. You can perform a medium change once a week, especially if the medium turns yellow after a few days. In principle, it is also possible to differentiate the cells with no further media changes.

  Related Links and Documents

  • CD34+ progenitor cells
  • Hematopoietic Progenitor Medium, ready-to-use
• To what extent will CD133+ Hematopoietic Progenitor Cells proliferate in Hematopoietic Progenitor Cell Expansion Medium XF (C-28021)?

  For optimal cell growth, human CD133⁺ Progenitors should be cultured in PromoCell Hematopoietic Progenitor Cell Expansion Medium XF (C-28021) supplemented with Cytokine Mix E (C-39890) or with any other cytokine cocktail suitable to achieve optimal cell expansion. Cytokine Mix E contains recombinant human TPO, SCF, flt3-ligand and IL-3. The strong expansion of the cells will persist for approx. 2 weeks. The expansion factor is typically 75-300-fold. Most of the resulting cells will however have lost the CD133 marker (CD133^–/CD38^–/CD34⁺) but they still represent primitive HPCs.

  Related Links and Documents

  • Hematopoietic Progenitor Cell Expansion Medium XF
• What markers does PromoCell test to characterize their Adult Human Stem and Blood Cells?

  All Adult Human Blood and Stem cells are analysed by flow cytometry to express defined markers. MSCs are additionally tested for their capacity to differentiate into the osteogenic, chondrogenic and adipogenic lineages. For more details, please see Certificate of Analysis of the respective cell type (“Phenotypic characterization”).

  Related Links and Documents

  • PromoCell Human Blood and Stem Cells
• What is the recommended split ratio for PromoCell CD34+ progenitor cells (C-12921)?

  PromoCell does not recommend a split ratio for human blood cells because the cell yield (number of cells per ml medium) will vary from lab to lab. Instead, we recommend to count the cells before subculture and calculate the respective split ratio by taking into account the recommended plating density. Please see the data sheet of hCD34⁺-CB cells or check the overview on our website for recommended plating densities.

  Related Links and Documents

  • Overview human stem and blood cells
• What happens if I grow CD34+ or CD133+ progenitors in Hematopoietic Progenitor Cell Expansion Medium XF (C-28021) without Cytokine Mix E (C-39890)?

  The cells will die. Hematopoietic Progenitor Cell Expansion Medium XF always must be supplemented with Cytokine Mix E or an appropriate cocktail of cytokines.

  Related Links and Documents

  • Human CD34+ Progenitor Cells
  • Hematopoietic Progenitor Cell Expansion Medium XF
• What is the difference between DC Generation Medium XF (C-28052) and DC Base Medium XF (C-28054)?
  • DC Base Medium XF (C-28054) consists of 250 ml Basal Medium + SupplementMix. It does not include cytokines and must therefore be supplemented according to the user’s individual needs.

  • DC Generation Medium XF (C-28052) is ready-to-use and consists of 250 ml Basal Medium + SupplementMix + appropriate cytokines.

  Both, Base Medium XF and Generation Medium XF have a xeno-free formulation.

  Related Links and Documents

  • DC Media XF
• What are the advantages of using PromoCell DC Generation Medium XF for dendritic cell generation?

  Our DC Generation Medium XF (C-28052) has a xeno-free formulation. It provides a complete media system (ready-to-use, all cytokines included) and shows efficient and reproducible in vitro maturation of moDCs from freshly isolated peripheral blood monocytes.

  Related Links and Documents

  • Dendritic Cell Generation Medium XF
• I would like to know if the Lymphocyte Separation Medium (C-44010) is endotoxin tested.

  Yes, our Lymphocyte Separation Medium 1077 is endotoxin-tested. The specification for the product is < 10EU/ml.

  Related Links and Documents

  • Lymphocyte Separation Medium
• How does PromoCell characterize their MSC-UC (C-12971)?

  Our hMSC-UC are characterized by their differentiation potential into chondrocytes, fat cells, and bone cells. In addition, we determine the presence of CD73, CD90 and CD105 expression as well as the absence of CD14, CD19, CD34, CD45 & HLA-DR expression by flow cytometry as proposed by the International Society for Cellular Therapy.

  Related Links and Documents

  • hMSC-UC
• What is the recommended storage temperature of PCR Mycoplasma Test Kit I/C (PK-CA91-1024) and I/RT (PK-CA91-3025C) ?

  The PCR Mycoplasma Test Kits I/C and I/RT should be stored at 4°C (lyophilized tubes). After rehydration, the recommended storage temperature is -20°C.

  Related Links and Documents

  • PCR Mycoplasma Test Kits
• Can you please let me know if the NAD/NADH Quantitation Kit (PK-CA577-K337) can be used on bacteria samples?

  We have optimized the assay protocol for samples of mammalian origin. However, theoretically it should work with samples from multiple origins including bacteria. Since we have not done it ourselves, I do not know if the assay protocol is the most optimized for such samples, so you may have to do some optimization to get the most efficient results. The one suggestion I can give is to increase the number of cells used, since bacterial cells are smaller than mammalian cells.

  Related Links and Documents

  • NAD/NADH Quantitation Kit
• I am having trouble with low signal or high background staining when using the Apoptotic/Necrotic Cells Detection Kit. Can I adjust the concentrations of Annexin V and Ethidium Homodimer III?

  The concentration of Annexin V and Ethidium Homodimer III can be increased or decreased if necessary to optimize staining for different cell types.

  Related Links and Documents

  • Apoptotic & Necrotic Cell Detection Kits
• Can I use the Apoptotic/Necrotic/Healthy Cells Detection Kit for staining live embryos, tissue explants, or organotypic slice cultures?

  The assays are designed to stain dissociated cells in culture and have not been validated for organ culture.
  Annexin V staining of early chicken and mammalian embryos in culture has been reported in the scientific literature. For staining of living tissues, the specimen would need to be thin enough to allow exposure of the cells to the 36 kDa Annexin V protein. Also, damage to cell membranes from dissection or sectioning of tissues could result in high background staining.

  Related Links and Documents

  • Apoptotic & Necrotic Cell Detection Kits