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

1989; Elsevier BV; Volume: 124; Issue: 2 Linguagem: Inglês

10.1016/0022-1759(89)90354-2

1872-7905

Naohiko Ikegaki, Roger H. Kennett,

Protein purification and stability

A simple method to increase the overall sensitivity of immunoblot assays is descrived. 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 ration 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.