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A general immunochemical method for detecting proteins on blots.

J M Kittler·N T Meisler·D Viceps-Madore·J A Cidlowski·J W Thanassi
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Abstract

Following horizontal electroelution, or blotting, of proteins from polyacrylamide gels to immobilizing matrices, such as nitrocellulose or Zeta-bind paper, the transferred proteins can be derivatized in situ with pyridoxal 5'-phosphate and sodium borohydride. After a quenching step to eliminate nonspecific binding of antibody to the protein-binding matrix, the blot is incubated with a solution containing a mouse monoclonal antibody specific for the 5'-phosphopyridoxyl group. The transferred proteins can then be located on the blot with second antibody staining procedures employing either a peroxidase-linked goat anti-mouse F(ab')2 antibody or a peroxidase-linked avidin/biotin system. The solid-phase enzyme-linked immunosorbent assay method described in this report is a mild, general, and sensitive immunochemical method for the detection of proteins on protein-binding matrices.

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AnimalsAntibodies, MonoclonalChromatinCollodionCytosolElectrophoresis, Polyacrylamide GelEnzyme-Linked Immunosorbent AssayHistonesImmunoenzyme TechniquesLiverNylonsPaperProteinsPyridoxal PhosphateRatsComparative StudyJournal ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, P.H.S.

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J M Kittler, N T Meisler, D Viceps-Madore, J A Cidlowski, J W Thanassi. 1984. A general immunochemical method for detecting proteins on blots.. https://doi.org/10.1016/0003-2697(84)90372-5

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