Hello,
I am using two different antibodies specific for the same membrane protein: one of them is monoclonal and the other polyclonal.
However, when I reveal the PDVF membrane the results are difficult to understand, for instance:
- when I use the monoclonal antibody my protein presents approximately 48 KDa; However, when I incubate with polyclonal one the same protein shows 35 KDa.
Do you have any idea of what might going on with this evidence?
Thank you for the attention.
To me it seems that only one of them is specific, but without looking at the actual gels and blots it is difficult to give a good answer! why don't you post some images of the blots?
Both antibodies are commercially available and specific for my target protein.
that doesn't mean anything, there are plenty of commercial antibodies that are specific according to the manufacturer but they end up not working or giving strange results, can you actually share the size of your protein, so that we know which antibody is the one working?
In your case, it is difficult to say what is the exact problem and below I would like to write them in increasing order of likelihood.
Without looking at your bands, it is impossible to say as the information is insufficient in your question. Also non of my answer will be valid if you get only one band (no duplicate bands together) with each antibody in the same sample. Do upload a photo of your bands.
Best,
Prabin
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