Journal
MOLECULAR AND CELLULAR PROBES
Volume 19, Issue 6, Pages 385-388Publisher
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
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Funding
- Intramural NIH HHS Funding Source: Medline
- NIA NIH HHS [Z01 AG000511-08] Funding Source: Medline
- 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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