If the protein A (60kda) and protein B (30kda) can form a protein complex together, is it possible to detect protein A band at 30kda (protein B location) when do western blot (incubating only anti-protein A)? Is SDS in Laminil's sample buffer sufficient to dissociate/solubilize all protein-protein interaction?
I have experienced a protein A detected at 30 kda but not sure whether it is really protein A, or left over protein band before stripping the membrane, or just another nonspecific band.
Thank you very much
Hi there,If the complex wasn't dissociated by the loading buffer, you would get a band heavier than 60kD. If you detect protein A in a 30kD band it's either a non specific signal or it's due to protein A degradation into a 30kD polypeptide retaining epitope recognized by the anti-protein A antibody.
I´m agree with Dominique. But you need to know if the protein A is a single or multimeric protein before attributing a possible hydrolysis of Protein A.
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