How to quantify protein which has sds buffer already - (Feb/19/2008 )
Hi,
I am running an ECL western blot using "drosophila head" protein as sample and try to compare the same protein from different lines. Since the sample got 0.1 % SDS when I extract the head. as a result, I can't quantify the protein using bradford method (gave me a incorrect quantification). So I give up this way and try to use a "guess" way to quantify my protein. for example,I use the same number of fly head (say 6 heads) to prepare the sample, however, even done this, still not able to get a even amount of protein for each samples.
Can anyone help about this? How to you quantify your protein for western blot?
Your kind help is appreciated!
Thanks
Hi Che,
I had a similar problem as I use RIPA buffer (also from Pierce) to lyse my samples which contains SDS. I use BCA assay from Pierce. Here is a link providing more info:
http://www.piercenet.com/Products/Browse.cfm?fldID=02020101
It is compatible with many substances and they have a table at the bottom of the page listing the different compounds that are compatible with the assay.
Good luck!