I use biotinylation kit to isolate various fractions from mammalian cell lysate. How to be sure what I see is membrane fraction. Also I need to see expression of my protein in the membrane fraction at high temperatures like 45 degrees. What is to be done to ensure good blot?
Taras Pasternak
Hi, Kalpita, you should find that marker which abundant only in the plasma membrane of your cell type. Indeed, it is difficult task, in many case you may have contamination with the other fraction, mainly inner cell membranes.
In any case it will be nice to confirm results with immuniocythochemistry. Good luck!.
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