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Skimmed‐milk blocking improves silver post‐intensification of peroxidase‐diaminobenzidine staining on nitrocellulose membrane in immunoblotting
Author(s) -
Ludány Andrea,
Gallyas Ferenc,
Gaszner Balázs,
Andrásfalvy Bertalan,
Szücs György,
Kellermayer Miklós
Publication year - 1993
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150140114
Subject(s) - nitrocellulose , immunostaining , staining , peroxidase , chemistry , skimmed milk , membrane , ascorbic acid , bovine serum albumin , chromatography , biochemistry , biology , enzyme , immunohistochemistry , food science , genetics , immunology
A silver‐staining procedure for enhancing the sensitivity of protein detection on nitrocellulose membranes in immunoblotting is described. After completing any peroxidase‐Ni‐diaminobenzidine immunostaining, nitrocellulose sheets are placed in a physical developer, containing sodium tungstate and ascorbic acid, until the desired grade of silver‐intensification has been reached. In this way a 16‐ to 64‐fold amplification of intensity of the initial immunostaining can be achieved. False positive silver staining of protein bands and of background are suppressed by replacing bovine serum albumin, the blocking agent most frequently used in immunoblotting, with skimmed milk.