My protein of interest gets detected in IF, but not by western. Protein is not expressed in cancer cells as the gene is carried the frameshift mutation. However, I saw ımmunofloroscence of the gene which targeted the same primer antibody and also I analyzed the mRNA expression of this gene. What's your suggestions?
hi
I agree with Sebastian . If you have enough Antibody , you can run IP test and so check in WB again .
good luck
Hi Gamze
I agree with Sebastian. You should check if your Ab is suitable for Western Blot. It might recognize a conformational epitope which means he woulnd't work for denaturated samples. You could try running a native PAGE and western blot, I believe it could work for conformational epitopes.
Yes it is possible, if your antibody recognize epitope which is far apart in amino acid chain but due to folding they become epitope.
So when you run SDS electrophoresis before western blotting the protein get denature and the epitope part get separated.
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