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Transfer of proteins from plastic‐backed isoelectric focusing gels to nitrocellulose paper
Author(s) -
Albaugh George P.,
Chandra G. Ram,
Bottino Paul J.
Publication year - 1987
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.1150080304
Subject(s) - nitrocellulose , electroblotting , isoelectric focusing , blot , collodion , chromatography , chemistry , antiserum , biochemistry , biology , antibody , membrane , immunology , enzyme , gene
Proteins were transferred to nitrocellulose paper from plastic‐backed isoelectric focusing gels. The technique involves a two‐step blotting procedure. In the first step, simple press blotting was used to establish adhesion between the gel and overlaying nitrocellulose paper. This step facilitated the removal of the plastic backing. In the second step, electroblotting produced a pair of bi‐directional replica blots. Incubation of the gel‐nitrocellulose paper sandwich in buffers containing T ween 20 permitted the intact release of the gel from the blots. The resulting blots were successfully stained with India ink or probed with antiserum.