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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