I'm differentiating hiPSCs into hepatocyte-like cells and I want to detect albumin by flow cytometry. I'm wondering what is the best protocol to do it. I read that for detection of secreted proteins it is good idea to use Brefaldin A to prevent protein release from the Golgi apparatus. Does anyone know anything about this?
Berfeldin A is anti-secretory, I worked with it to prevent secretion of IFN- alpha by PBMC, but it could be used to prevent ER-Glogi transport in general.
For some proteins Brefeldin inhibits better and for some monensin inhibits better. Sometimes the combination also inhibits well. I suggest you to check which inhibitor works better for albumin. Either brefeldin or monensin or both in combination.
The papers I found online looking at intracellular albumin expression have done so without using any of the secretion inhibitors. If you think that would help, try using a mix of BFA and monensin like Pragun Muthya suggested. However, keep in mind that the longer you incubate the cells, the more they die.
https://www.pnas.org/content/early/2017/03/21/1618291114#sec-9
Article Enhanced Differentiation of Adult Bone Marrow-Derived Stem C...
https://bitesizebio.com/36433/brefeldin-monensin-hunt-proteins/
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