Protein quantification using HPLC - (Mar/27/2013 )
Does anyone have a good reference w/ a protocol for using HPLC and a standard curve to quantify protein levels in a cell lysate? My antibody seems to be terrible for performing a western blot, I wonder if it will work better to IP out the protein then detect it using HPLC? I can buy the protein in question from a vendor, and assume just make a standard curve, measure peak area via HPLC, come up with a concentration standard curve.
that should work. the software that comes with some hplcs allow you to identify and quantitate the peaks. just make sure that the peaks are well separated and reproducible.