I am recently doing the EMSA for the AhR and related binding sequence probe.
We use the rat liver nuclear extract done by a commercial kit. Pierce EMSA lightshift kit was used
I can not big difference of bind with cold competitor ( non labeled probe same as the biotin labeled probe). Then I added a consensus XRE probe which is supposed to be bound by AhR to compete the labeled probe,I failed.
I am just wondering if none of the competition works, does that mean the binding shift we found is non specific?
free probe worked and kit quality controls worked.
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