I would like to check the expression profile of some proteins of PBMC and PBMC derived macrophages(with treatments). For the same purpose, what would be a good HKG for western blot? Any of your experiences are welcome!
I have never work with those cell types, but a general rule is to validate tubulin, actin and GAPDH: run a WB with the same amount od protein for all the samples you have (PBMC and macrophages), do a WB with anti-tubulin, another with anti-actin and another with anti-GAPDH antibody and check which protein has the same expression in all your samples. You can then use that protein as a reference for relative quantification.
Yap you are right, it is worth to make a test of all HKG with both cell types with treatments and find out the better one :)
Depending on what you do with the cells (Knock-down of genes may affect your potential house-keeping genes) you have to test a number of them to find a stable one.
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