4.4 Article

Clean western blot signals from immunoprecipitated samples

Journal

MOLECULAR AND CELLULAR PROBES
Volume 19, Issue 6, Pages 385-388

Publisher

ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.mcp.2005.06.007

Keywords

heavy Ig chain; light Ig chain; protein A-HRP; protein-G-HRP; background western signals

Funding

  1. Intramural NIH HHS Funding Source: Medline
  2. NIA NIH HHS [Z01 AG000511-08] Funding Source: Medline
  3. NIGMS NIH HHS [R01 GM62803, R01 GM062803] Funding Source: Medline

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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. (C) 2005 Elsevier Ltd. All rights reserved.

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