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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.

Struggling with faint or invisible bands in your Western blots? This in-depth guide provides a detailed, step-by-step protocol for detecting low-abundance proteins. We cover critical optimizations from strategic sample preparation and lysate concentration to advanced detection using high-sensitivity ECL substrates. Learn how to choose the right buffers, membranes, and antibodies, and master techniques for enhancing your signal-to-noise ratio.