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Problem in Bradford assay

Problem in Bradford assay - Protocols and Methods Forum

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  #1  
Old 08-21-2009, 03:40 AM
Pipette Filler
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Default Problem in Bradford assay



I tested protein level of my microsome samples with Bradford assay. I did serial dilution of my samples and than found the original concentration of my samples.

I found that there were large difference in original concentration calculated from samples with different dilution factors.

In my sense, dilution factor must not affect the calculation of original concentration of a substance. But it happened. Can anyone help me???

Thx!!!
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Old 09-18-2009, 07:45 AM
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Default Re: Problem in Bradford assay

I had the same problem with my protein measurement but I do not know why with different dilution factor, the calculated protein concentration varies. Can someone please explain?
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Old 09-18-2009, 09:01 AM
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Default Re: Problem in Bradford assay

Bradford assay is quite sensitive to detergents and other things that might be in lysis buffers. What is the buffer the microsome samples are in? What is your dilution buffer?
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Old 09-25-2009, 09:22 AM
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Default Re: Problem in Bradford assay

My protein sample is dissolved and kept in TES buffer (Tris,EDTA,Sucrose). In the Bradford assay, I only use sterile distilled water to dilute my protein sample. In this case, is there alternative to avoid this problem other than changing the protein storage buffer (TES)?
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