HI, I am working on nfkb1 and I need to check its expression on Raw and THP-1 cells by western as well I need to immunoprecipitation for its interacting partners.
I have taken p50 ab from e-biosciences, this is working well with total lysate but when I am checking expression on nuclear and cytoplasmic fractions, I am not getting proper ab binding, Is this make difference.
When I did immunoprecipitation with p50, its giving very weak band , so I wonder how should i look for its intercating partners.
I am planning to order new antibody, could anyone suggest me if somebody has already working on same portein and cell lines?
ECM Biosciences has a rabbit pAb that they say will blot mouse NFkB in RAW. Here is our listing of their Ab: https://www.linscottsdirectory.com/antibodies/ecm-biosciences-llc/NFkappaB-p65-9666596
Please contact them directly for more details.
Shilpi, try once again with more number of cells for nuclear-cytoplasmic fractions...as Ab is working for immunoprecipitation. There might be some issue with your protocol.....or number of cells...
All the best...
Shilpi before switching to a new antibody, you can try with increasing amount of protein.
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