BCA Assay and SDS-PAGE Gels Do Not Agree (View forum version)


Posted 18 August 2010 - 12:11 PM

Hello All.

I am running a protein on a BCA Protein Assay (Pierce) and getting good results. I loaded 20 uL of the undiluted sample (approx 2ug/uL according to BCA, so 40 ug)on a denaturing 4-20% Novex SDS-PAGE gel with 5 uL sample buffer and see absolutely nothing. Any suggestions/insight would be greatly appreciated.


Posted 18 August 2010 - 07:15 PM

I dunno what concentration sample buffer you are using, but usually I use a 1:1 volume ratio of sample buffer to diluted/undiluted sample

And what do you mean by "See absolutely nothing"? You mean you don't see the protein lanes running down or something?


Posted 19 August 2010 - 05:20 AM

Have you fixed your gel before staining? What kind of staining have you used?