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Updated: Aug 16, 2026

Western Blotting: Sample Preparation to Detection
Published on: October 14, 2010
Clean Western blot signals from immunoprecipitated samples
Ashish Lal1, Susan R Haynes, Myriam Gorospe
1Laboratory of Cellular and Molecular Biology, National Institute on Aging-IRP, National Institutes of Health, Box 12, 5600 Nathan Shock Dr., Baltimore, MD 21224, USA. ashishl@grc.nia.nih.gov
Abstract:
We present a strategy that overcomes the high background arising during Western blotting (WB) detection of proteins obtained through immunoprecipitation (IP). Traditional HRP-conjugated secondary antibodies, which detect the denatured heavy and light antibody chains, produce high background that often mask the signals of interest on WBs. Here, we show that HRP-conjugated Protein A and Protein G, which detect almost exclusively intact antibody molecules, can be effectively used to obtain clean and specific WB signals of target proteins.
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