I'm doing a Bradfod assay but currently I'm still confused as to what blank i should use for my sample, which is the supernatant of PAO1 culture grown in AB minimal medium. For the standard, i use serially diluted BSA in dH2O, so the blanks should be a mixture of Bradford reagent and dH2O. But for the sample's blank, what should i use? Should i use the dH2O blank as well, but also add another sample of fresh AB medium, then subtract the absorption of fresh AB medium from the absorption of PAO1 culture's supernatant??
for blank reading. use all reagents which used for measure bradford assay without sample
Hi @Muh Risky
You should use fresh AB medium instead of dH2O, as blank for your test protein quantification.
What u are doing is right. But would also prepare my bsa standards in ab medium. Ideally I would go for precipitation and dissolve the protein in appropriate buffers.. And use this buffer as blank as well as preparing my bsa standards.
for blank reading. use all reagents which used for measure bradford assay without sample
You should consider blank as a solvent which you use to dissolve BSA/ or protein etc..
So Blank should be your solvent only ( it may be PBS, NS or distilled water).
Hi Muh,
I think Bradford aasay is not that much sensitive excercise.
You can use distilled water as blank.
Thank you
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