How to measure total protein conc. of cultured-cell lysate?

Mohamed BaSalamah mbasal at med.unc.edu
Mon Dec 9 19:18:40 EST 1996


Hellow

I wanted to thank all those who responded--privatly and on this
beautiful newsgroup--to my help request.

In addition, I used the BCA method from Pierce successfully. 

I have to make BSA standards in the same buffer solution and do 5 folds
dilution. That is taking the SDS concentration to 0.4% in all sample and
standards.

I added the other components of the SDS gel loading buffer (DTT and
BrPhenol Blue) after I'm satisfied with the measurments.

----- From my small experience, direct UV measurment is not reliable,
the presence of 1% SDS does not allow direct total protein measurment
using Peirce BCA's nor Biorad's kits.

Mohammed BaSalamah



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