Protein quantification after 8M urea lysis - (May/10/2011 )
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.
you can dilute your sample
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!!!
or you can do drop dialysis but you may lose some protein if it binds to the dialysis membrane.