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This frustrating (and common) problem can ruin days of work, but the fix is often simple. Our comprehensive troubleshooting guide diagnoses the real reason for your patchy bands. We cover everything from the #1 culprit—air bubbles in your transfer sandwich—to aggregated antibodies, uneven gel polymerization, and improper sample prep. Stop guessing and start fixing. This article provides clear, actionable solutions to rescue your blot and get the clean, sharp bands you need.

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.