Glutaraldehyde fixation of the primary antibody-antigen complex on nitrocellulose paper increases the overall sensitivity of immunoblot assay.
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.
Ikegaki
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N
Department of Human Genetics, University of Pennsylvania, School of Medicine, Philadelphia 19104.
Kennett
R H
RH
D000447:Aldehydes; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D015153:Blotting, Western / Q000379:methods*; D003101:Collodion; D004591:Electrophoresis, Polyacrylamide Gel; D005404:Fixatives; D005976:Glutaral; D007124:Immunoenzyme Techniques; D016274:Oncogene Protein p55(v-myc); D015203:Reproducibility of Results; D015682:Retroviridae Proteins, Oncogenic / Q000276:immunology; D014407:Tumor Cells, Cultured
D000447:Aldehydes; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D005404:Fixatives; D016274:Oncogene Protein p55(v-myc); D015682:Retroviridae Proteins, Oncogenic; D003101:Collodion; D005976:Glutaral