Protocol Online logo
Top : New Forum Archives (2009-): : Protein and Proteomics

Too bright blue for bradford assay - (Jul/16/2009 )

Hi. I tried to maesure protein conc. by Bradford assay.
I got satisfactory standard curve, but I found something wierd.
My sample with bradford reagent is too bright. Actually I found the mixture had highest absorbance at 625nm.

What's wrong with my sample? Actually I avoided using detergents.
Thanks in advance.

-yja97-

yja97 on Jul 16 2009, 10:20 AM said:

Hi. I tried to maesure protein conc. by Bradford assay.
I got satisfactory standard curve, but I found something wierd.
My sample with bradford reagent is too bright. Actually I found the mixture had highest absorbance at 625nm.

What's wrong with my sample? Actually I avoided using detergents.
Thanks in advance.



Could it be too concentrated? Try diluting it 1 in 10 or more.

-fishdoc-

Thanks for reply, fishdoc.
But I don't think my protein has high conc. I'm expecting around 10 ug/ml.

-yja97-

What else are in the buffer? Detergent components, such as Triton X 100 can interfere the results.

-genehunter-