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We are interested in your antibody to phospho-P21. Wonder why the band of phospho-P21 on Western Blotting is located between 26K and 36K. It is highly expected to be located around 20K. I am looking forward to your suggestions.
Asked on Sep 12 2007
Thank you for your enquiry. The reason why the detected and expected bands are different is maybe due to modifications like the following: 1) post-translational modification - e.g. phosphorylation, glycosylation etc which increases the size of the protein 2) post-translation cleavage - e.g. many proteins are synthesized as pro-proteins, and then cleaved to give the active form, e.g. pro-caspases 3) splice variants - alternative splicing may create different sized proteins from the same gene 4) relative charge - the composition of amino acids (charged vs non-charged) multimers - e.g. dimerisation of a protein. This is usually prevented in reducing conditions, although strong interactions can result in the appearance of higher bands I am afraid that we do not know the exact cause of the difference in band size. The image that we have was the outcome our laboratory obtained during their testing. I am sorry I can't be of more assistance in this occasion but should you require any other information, please do not hesitate to contact me.
Answered on Sep 12 2007