Premium
Direct immunoblotting from histological sections of brain onto nitrocellulose: Evaluation with the protein neurophysin
Author(s) -
Reisinger Peter W. M.,
Unger Jürgen W.
Publication year - 1990
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.1150110809
Subject(s) - neurophysins , nitrocellulose , collodion , antibody , membrane , chemistry , biochemistry , peptide , microbiology and biotechnology , biology , immunology , hormone
A method for the isolation and localization of proteins and peptides from histological sections of rat and human brain by immunoblottin is described. For validation, the well‐characterized protein neurophysin was electrophoretically transferred from formaldehyde‐fixed or fresh tissue sections onto a nitrocellulose membrane. Neurophysin on the nitrocellulose membrane was detected by a specific antibody reaction. The antibody against neurophysin was visualized either by using secondary antibodies, conjugated with peroxidase or by protein A gold, followed by enhancement with silver. With this simple and fast method, neurophysin (or other proteins and peptides) can be identified on nitrocellulose membranes in areas that correspond to anatomically defined regions. Since the procedure combines the advantages of precise regional localization of polypeptides with the specificity of antibody‐antigen reactions, the method may prove useful for rapid screening of the distribution of peptides or proteins in (brain) tissue.