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Can I replace HSA with FBS in the Macrophage Detachment protocol? Why is 2 mM EDTA added to the PBS?
In the detachment protocol for PromoCell’s Macrophage Detachment Solution, Human Serum Albumin (HSA) is recommended as an additive to PBS.
Use of FBS instead of HSA- FBS (or BSA) can technically be used as a substitute for HSA.
- However, this is not recommended, as FBS can cause unwanted immunological stimulation of human macrophages.
- Such stimulation may alter macrophage activation status, gene expression, or downstream functional readouts.
For this reason, HSA is preferred to maintain macrophages in a more defined and physiologically neutral state during detachment.
Role of EDTA (2 mM) in the PBS - EDTA chelates Ca²⁺ and Mg²⁺ ions, which are required for many cell–cell and cell–surface adhesion mechanisms.
- In combination with Ca²⁺/Mg²⁺‑free PBS, EDTA:
- Further reduces cell–cell adhesion
- Enhances the anti‑clumping effect
- Facilitates gentle detachment of macrophages without enzymatic treatment
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