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Bradford assay blank....why bother?? - (Jul/30/2012 )

Im just reading the procedure for a biorad (bradford assay) which involves the preparation of a protein blank which is subtracted from all standards and the sample?

Why bother, surely the blankis the same for all of them (unless one has made up a sample in a completely different buffer) so if one reads of the standard curve, one would get the same answer???

Am I wrong?

Confused.com

-johnuknow-

you need the blank to zero the spectrophotometer.

-mdfenko-

you need the blank to zero the spectrophotometer.

But if I plot the standards without zeroing against then read off the sample tube (again without zeroing).....I will get the same answer as if I had zero. Essentially what zeroing does is subtract the same value from exerything...ie standards, samples etc etc.

-johnuknow-

blanks for the bradford are relatively high absorbance. if you don't zero then you may be taking readings beyond the linear range of the spectrophotometer.

-mdfenko-