I’ve been having weird background signals even after trying different blocking buffers. Could it be the antibody itself causing nonspecific binding? If so, how do you choose a more specific one that actually gives clean bands?
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Blocking gave me headaches for months — I tried everything from milk to BSA, and the results were always inconsistent. Eventually, I realized it wasn’t just the buffer; washing steps were my real issue. Rushing through washes leaves background behind. Since then, I started timing each wash carefully, and suddenly my blots looked professional. Sometimes, it’s not about new reagents — it’s about discipline.