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Too bright blue for bradford assay


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3 replies to this topic

#1 yja97

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Posted 16 July 2009 - 07:20 AM

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.

#2 fishdoc

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Posted 16 July 2009 - 07:38 AM

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.

#3 yja97

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Posted 16 July 2009 - 10:30 AM

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

#4 genehunter

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Posted 16 July 2009 - 10:33 AM

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




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