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Characterization of a panel of neurofilament antibodies recognizing N‐terminal epitopes
Author(s) -
Kaplan M. P.,
Chin S. S. M.,
Macioce P.,
Srinawasan J.,
Hashim G.,
Liem R. K. H.
Publication year - 1991
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.490300312
Subject(s) - neurofilament , polyclonal antibodies , keyhole limpet hemocyanin , epitope , microbiology and biotechnology , vimentin , monoclonal antibody , antibody , glial fibrillary acidic protein , immunofluorescence , transfection , biology , blot , cytoskeleton , chemistry , immunohistochemistry , cell culture , biochemistry , immunology , gene , cell , genetics
Peptides corresponding to sequences from the ammoterminal “head” regions of the low, middle, and high molecular weight neurofilament proteins (NF‐L, NF‐M, and NF‐H) were synthesized by a modification of the Merrifield solid‐phase method, and a panel of polyclonal antibodies to these epitopes were prepared in rabbits by the injection of synthetic peptides conjugated to the carrier protein keyhole limpet hemocyanin (KLH). An additional, monoclonal antibody recognizing both glial fibrillary acidic protein (GFAP) and vimentin was also produced, by fusion of cells of the mouse myeloma line NS‐1 with spleen cells from a mouse immunized with cytoskeletal extracts. Antibody specificities were confirmed by a combination of Western blotting against cytoskeletal extracts and immunofluorescence using both rat brain sections and fibroblasts transfected with fully encoding cDNAs for each neurofilament protein, driven by viral promoters.