I have to elute my protein in Sucrose buffer and I wondering how accurate would bradford assay be? Would it react to the Sucrose thats inside the buffer (250mM)?
Thank
I am not sure what kit you are using but according to the kit on thermo's website the assay should be able to handle 10% sucrose (290mM) therefore I think this should be ok.
Kit link:
https://www.thermofisher.com/order/catalog/product/23200
Bradford assay can detect low concentration proteins up to 1 ug/ml. I'm not sure sucrose can bind with CBB dye.
I would suggest that if you have Bradford reagent available to you that you construct a standard curve using the normal process exept using samples prepared with the appropriate amount of sucrose. You can compare that with standards prepared following the instructions that come with the reagent. Compare the plots for differences in slope and linearity in the range you expect to use the Bradford method. If you decide to use the Bradford reagent with your samples. Construct the standard curve usng the sucrose buffer.
Also, prepare blanks containing only sucrose & your other reagents, minus protein, to see if sucrose contributes anything to the color reaction.
Yes, use your sucrose buffer just as you would with your aexperimental samples. Good idea to make some blanks as James Bonitati says. This would test for interference (background from the sucrose - (if there were some impurities that would react with the Bradford reagent).
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