if i made ten percent brain homogenate and i want to add lysis buffer suppose RIPA buffer...what should be the ratio of PBS made homogenate to RIPA buffer?
thanks
RIPA seems rather harsh - are you trying to preserve native proteins? When I generate brain lysates I use cold MES buffer and dounce homogenization. If you know for certain RIPA is ideal for your application and you have a stock of brain lysate already and need to generate at 10% dilution with PBS, you'd take, for example, 1 mL of lysate and dilute with 9 mL PBS.
thanks for your answer and i am bit new to the neuroscience field so not sure which buffer would better suit me...actually my target protein in first instance is prion protein scrapie and later on i would like to detect CREB and BAD proteins both in total and phosphorylated state...thanks again SIr...
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