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Protein quantification after 8M urea lysis


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#1 NTL

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Posted 10 May 2011 - 10:16 AM

Hi,
I used 8M urea (+ phosphatases) to lyse cells. I am not sure how to measure the protein concentration after lysis. BioRad protein assay say we can measure the protein concentration in urea up to 6M only. Do I need to use the DC BioRad assay for measurement but the mannual says this kit's reagents are compatible with up to 4M urea? How low can the urea concentration be made and we still can lyse the cells? Thank you very much for your help.

#2 mdfenko

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Posted 10 May 2011 - 10:28 AM

you can dilute your sample
talent does what it can
genius does what it must
i do what i get paid to do

#3 Prep!

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Posted 10 May 2011 - 07:28 PM

yeah unless u think ur protein is in very low concentration which cant be detected after dilution... dilution is the simplest option available here!!!
alternatively u can buffer exchange or dialyze but the be ready to account for losses and validate the sample preparation step!!!
Support bacteria - They are the only culture some people have!!!
Cheers!!!

#4 mdfenko

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Posted 11 May 2011 - 08:20 AM

or you can do drop dialysis but you may lose some protein if it binds to the dialysis membrane.
talent does what it can
genius does what it must
i do what i get paid to do




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