I want to determine the protein concentration of a self isolated S9 fraction from a cow liver using a Bradford assay. It is however unclear to me if I should use a lysis buffer before performing a Bradford assay (most protocols I find online are performed using cell culture instead of cellular fractions). If I need to use a lysis buffer will Triton X-100 work and in what quantities should I use it? (How many mL lysis buffer per mL S9 fraction?
I hope someone can help me
Yes you should lyse the cells first. Snap freezing and mechanical homogenization can work. Sonication is also an option. Use 0.5% Triton X-100.
An S9 fraction is prepared from cells that have already been lysed, so there should not be any need to use a detergent prior to running a Bradford assay. In fact, detergents interfere with the Bradford assay, so it would be better not to use one.
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