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[DOI] [PMC free article] [PubMed] [Google Scholar] [108].Costa MC, Silva G, Ramos RV, Staempfli HR, Arroyo LG, Kim P, et al

Mao et al., 1999

Assuming you followed the above advice and used an appropriate binding buffer and your protein isnt huge, then a likely reason for this is that the expression yield is much lower than you expected

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p53-pS15 is a major focal point in p53 activation to mediate p53-dependent transcription or growth arrest, Ser 15 phosphorylation triggers a sequence of additional sites phosphorylation in p53 (including Ser9, -20, -46 and Thr18) that contribute further to p53 activation 28