I want to measure protein levels of cells attached to a 96 well plate. If the lysis buffer is added to the plate itself, is it alright if the cell debris remain in the plate (if any remain after lysis) and bradford reagent is directly added into the wells? Or is it necessary to centrifuge out the cells after addition of lysis buffer to the wells?
The Bradford assay is an absorbance assay. If there is any debris in the well that is not dissolved by the lysis buffer or dye reagent, it will scatter light and give a falsely elevated, and probably noisy signal. A clear solution is required.
I agree with adam, we use to collect lysate and transfer them into a v-bottom 96 well plate to carry out centrifugation and used the clear lysate to measure the protein conc.
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