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Western Blot Woes: Why Is My Protein Running at the Wrong Molecular Weight?
Is your Western Blot showing a band at the wrong molecular weight? This is one of the most common frustrations in the lab, but it's not a dead end. An unexpected band size is often a critical clue, not a failed experiment. The theoretical calculated weight from amino acid sequence rarely matches the observed weight on a gel. This discrepancy can be caused by a host of biological reasons from post-translational modifications like glycosylation to protein cleavage, splice varia
Dec 10, 20258 min read
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Antibody Not Working for Western Blot? Guide to Troubleshooting No Signals
When your antibody fails to detect a signal, the problem can be complex. Is it the antibody itself? The sample? Or the protocol? This guide provides a complete troubleshooting protocol for "no signal" Westerns. We dive into the most common reason for failure and then provide a systematic checklist. Learn how to diagnose issues with your transfer using Ponceau S, why your blocking buffer might be the culprit, and how one common preservative could be killing your signal.
Dec 5, 20255 min read
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