I have treated the cells with complex and after a period of time I am isolating the protein for expression studies. During this step should I isolate protein from live cells alone or both the cells. If both the cells, should I isolate together or separately? Then how to interpret the results?
Fisher Scientific has a number of kits available for protein collection, which I use often. I've used both their nuclear and cytoplasmic kits as well as the subcellular fractionation kits which gives good yield of protein. If using adherent cells in your model, trypsinize the cells, spin down and lyse per the protocol (make sure you have both protease and phosphatase inhibitors!).
Please clear your question, what do you mean complex(some factors? ok) and what are both cells? (treated and untreated?)
Both the cells means live cells and dead cells. Metal Complex used for treatment like cisplatin. After treating the cells with Ic50 concentration i am going for protein isolation from the treated cells. In this stage shall i discard the dead cells or should i isolate protein from both live and dead cells.
Hello there,
I understand that you are interested to study the effects of metal complex on your cells. After treating the cells with a certain amount of the complexes, generally we would like to know the expression variation in which the live cells are responding to. This would mean to discard the dead cells and isolate the protein from the live cells. Hope this helps. Good luck !
Regards,
Naresh
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