Glutaraldehyde fixation of the primary antibody-antigen complex on nitrocellulose paper increases the overall sensitivity of immunoblot assay

J Immunol Methods. 1989 Nov 30;124(2):205-10. doi: 10.1016/0022-1759(89)90354-2.


A simple method to increase the overall sensitivity of immunoblot assays is described. The method is based on fixation of the primary monoclonal antibody-antigen complex on nitrocellulose paper by a divalent crosslinker, glutaraldehyde. The fixation of the antibody-antigen complex significantly increases the sensitivity of the assay. Unexpectedly, the glutaraldehyde treatment also improves the signal/noise ratio by decreasing non-specific binding of secondary antibodies and/or tertiary reagents. Thus, a significant overall improvement of immunoblot assays is achieved by this method.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Aldehydes*
  • Antibodies, Monoclonal
  • Antigen-Antibody Complex*
  • Blotting, Western / methods*
  • Collodion*
  • Electrophoresis, Polyacrylamide Gel
  • Fixatives*
  • Glutaral*
  • Immunoenzyme Techniques
  • Oncogene Protein p55(v-myc)
  • Reproducibility of Results
  • Retroviridae Proteins, Oncogenic / immunology
  • Tumor Cells, Cultured


  • Aldehydes
  • Antibodies, Monoclonal
  • Antigen-Antibody Complex
  • Fixatives
  • Oncogene Protein p55(v-myc)
  • Retroviridae Proteins, Oncogenic
  • Collodion
  • Glutaral