I have used a transcription factor (pure protein) which is 80kDa and got a binding to a specific sequence at 1ul. Braddford conc. of total protein was 1.6ug/ul and for the specific protein band around 0.5 ug/ul after deduction. So, is the binding stong enough . Carrier DNA was used at .25 ug/ul (polydI/dC). Secondly, how polydI/dC acts as competitor DNA. How does it reduce non specific background?
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