Clean Western blot signals from immunoprecipitated samples

Mol Cell Probes. 2005 Dec;19(6):385-8. doi: 10.1016/j.mcp.2005.06.007.


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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, N.I.H., Intramural

MeSH terms

  • Blotting, Western / methods*
  • Horseradish Peroxidase / chemistry
  • Immunoglobulin Heavy Chains / chemistry
  • Immunoglobulin Heavy Chains / metabolism
  • Immunoglobulin Light Chains / chemistry
  • Immunoglobulin Light Chains / metabolism
  • Immunoprecipitation / methods*
  • Nerve Tissue Proteins / chemistry
  • Nerve Tissue Proteins / metabolism
  • Protein Binding
  • Staphylococcal Protein A / chemistry
  • Staphylococcal Protein A / metabolism


  • G-substrate
  • Immunoglobulin Heavy Chains
  • Immunoglobulin Light Chains
  • Nerve Tissue Proteins
  • Staphylococcal Protein A
  • Horseradish Peroxidase