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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.
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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.
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You may find all information regarding the activation and the cytokine concentrations in table 1 (page 5) of our Application Note.
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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.
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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.
Further Information
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
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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.
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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.
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Yes, it is possible to refreeze them.
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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.
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The PBS buffering enhances the Alizarin Red staining (precipitation of the dye) and makes it more intense. Leave the PBS on the cells after staining/washing and analyze the sample immediately, as the dye may bleed upon prolonged storage without embedding.