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Protein blotting analysis of large sample numbers
Author(s) -
Kapfhammer Josef P.
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.11501401140
Subject(s) - blot , monoclonal antibody , antibody , microbiology and biotechnology , chemistry , chromatography , biology , biochemistry , immunology , gene
A method is described that allows probing of hundreds of different antibodies on protein blots per day, making it possible to use protein blots for primary screening of hybridoma supernatants. It involves a miniaturization of the protein blots and their attachment to a solid support. This allows a great reduction of the required number of gels and electroblots and brings a drastic simplification of the incubation and washing procedures. Apart from a reduced resolution of closely adjacent bands the resulting mini‐blots are comparable to conventional protein blots in sensitivity and specificity. The procedure has been successfully applied for primary screening of hybridoma supernatants. It can be useful in cases where a monoclonal antibody to an antigen with known molecular weight is required, or in situations where many different antibodies have to be tested in a short time period.