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Polyclonal Antisera to the Individual Neurofilament Triplet Protdins: A Characterization Using ELISA and Immunoblotting
Author(s) -
Karlsson J.E.,
Rosengren L. E.,
Haglid K. G.
Publication year - 1989
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1989.tb11770.x
Subject(s) - antiserum , neurofilament , polyclonal antibodies , kilodalton , microbiology and biotechnology , protein subunit , biology , antigen , biochemistry , immunohistochemistry , immunology , gene
Abstract In this article, the preparation and characterization of polyclonal rabbit antisera against the individual polypeptides of bovine neurofilament (68, 150, and 200 kilodaltons) is described. Selected antisera against the 68‐ add 150‐kilo‐dalton neurofilament polypeptides were specific for the corresponding antigen in homogenates of bovine, rat, and human brain as judged by immunoblots. The antisera against the 200‐kilodalton neurofilament polypeptide crossjreacted to some extent with the 150‐kilodalton neurofilament polypeptide, especially with the human antigen. The most specific antisera were used to develop an enzyme‐linked immunosorbent assay (ELISA), and the cross‐reactivities between the antisera and the different bovine and rat neurofilament polypeptides were determined. Contrary to the results In the immunoblots, the antiserum against the 200‐kilodalton neurofilament polypeptide was subunit‐specific, as was the 150‐kilodalton antiserum. The 68‐kilodalton antiserum displayed a minute cross‐reactivity against bovine 150‐ and 200‐kilodalton neurofilaments, but it cross‐reacted somewhat more with the rat 150‐ and 200‐kilodalton antigens. Even so, the subunit specificity of the antisera is high enough to enable the development of a quantitative ELISA for determination of the individual bovine or rat neurofilament polypeptides in a mixture. This study is the necessary preparation for such an assay.