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Protein quantification using HPLC


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#1 Ubiquitous

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Posted 27 March 2013 - 10:53 AM

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.

#2 mdfenko

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Posted 28 March 2013 - 04:45 AM

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.
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