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protease inhibitors and bradford


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#1 jasmina

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Posted 17 November 2010 - 04:11 AM

Hi all,
I ve qantified protein extract of brain tissue, solubilized in NP40buffer with protease inhibitors, phosphatase inhibitors and sodium floride.
so, im wondering if this could interfere with bradford and affect the quantification (BSA). also i use blank in which i put the buffer NP40 with that inhibitors as well.
thanks for advice

#2 mdfenko

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Posted 17 November 2010 - 08:40 AM

many protease inhibitors are peptides and will interfere with protein assays. using a blank containing the inhibitors at the same concentration as the samples will help, especially if the sample protein is in greater abundance than the inhibitors.

but, the readings must remain within the range of the standards (you can also add the blanking buffer to the standard curve).

Edited by mdfenko, 17 November 2010 - 08:42 AM.

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i do what i get paid to do

#3 jasmina

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Posted 17 November 2010 - 02:18 PM

ok!! of course, the blank is added to the BSA curve
thanks




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