PROTEIN ASSAY - MEASUREMENT OF PROTEIN IN CELL LYSATES (May/23/2002 )
I need to measure total concentration of protein in a cell lysate. Can anyone help me?
You can try protein estimation by standard protocols using reagents like bradford's , Lowry, BCA.Protocols are available in any standard protein protocol books. You may choose the assay system depending on the compatibility of your bufffer with the assay reagent. For every reagent there are certain components that interfere with the assay system. "Methods in Enzymology vol. 182"pp.50-69 gives a very good idea of what assay one could use for a particular system as it has a table is which gives an overview of various assay procedures and corresponding substances that interfere with the assay.
" Current protocols in protein science" also gives a good idea of various methods for protein estimation.
A BioRad Bradford-based protein assay reagent has always worked well for me. If your lysate contains fair amount of detergent, there is DC (detergent compatible) version of it. Also, Pierce offers BCA protein assay reagent.