Aside from re-reunning your gel with samples correctly prepared, probably not I'm afraid. It's probable that your primary antibody only recognises a reduced form of the epitope (the beta-mercaptoethanol reduces disulphide bonds in the loaded proteins), and therefore it will not bind as efficiently or will not bind at all - if you loaded reduced samples next to non-reduced samples it is likely that diffusion of the B-ME may have partially reduced those non-reduced samples. If you can find an alternative antibody which recognises a non-reduced epitope, then you might be able to get a result with it after stripping the already bound antibodies off the membrane (there are protocols readily available online if you don't know how to do this). Antibodies which say they can be used for ICC or flow cytometry might be better to look for, as they will almost certainly recognise a non-reduced epitope.

I did think about whether you could try reducing the proteins bound on the membrane by soaking in a reducing agent, but a person here tried that and said it didn't work for them. https://www.researchgate.net/post/Is_it_possible_to_reduce_the_protein_already_in_a_western_blot

Hi Anika Sanjana

I agree with Samuel Davis

you can check if the antibody epitope can recognize your protein in nonreduced form. if not then I am afraid you have to add B-ME to samples if you still have any left. or you will need to prepare new samples.

But by the way did your antibody give any signal with the reduced ones?

i do have samples left however, even after reboiling them with BME it did not show any signal. I can try a different antibody.