TY - JOUR
T1 - Domain structure of neurofilament subunits as revealed by monoclonal antibodies
AU - Angeletti, Ruth Hogue
AU - Trojanowski, John Q.
AU - Garden, Martin
AU - Schlaepfer, William W.
AU - Lee, Virginia M.‐Y
PY - 1985
Y1 - 1985
N2 - Monoclonal antibodies have been prepared against purified neurofilament (NF) subunits (NF68, NF150, and NF200). From 25 fusions, several hundred strongly positive antibodies have been obtained. Among them are antibodies against the specific subunits as well as antibodies recognizing common antigenic determinants. These have all been characterized according to the following properties: ELISA (enzyme‐linked immunosorbant assay) testing against each subunit, immunoblots against enriched neurofilament preparation, immunoblots of cyanogen bromide or chymotrypsin‐treated neurofilaments, immunofluorescence with PC12 cells, and immunohistochemistry of cerebellum. Whereas the antibodies against the NF68 and NF150 appear to react with single cyanogen bromide fragments, the antibodies against the NF200 react with multiple cyanogen bromide fragments. These data are consistent with the hypothesis that the NF200 is partially computed of several repeated structural determinants. Furthermore, all of the antibodies that react with the NF200 recognize the solubilized “sidearm” domain from limited chymotryptic digestions. The locations of the common and variable domains of the three subunits are discussed in light of these results.
AB - Monoclonal antibodies have been prepared against purified neurofilament (NF) subunits (NF68, NF150, and NF200). From 25 fusions, several hundred strongly positive antibodies have been obtained. Among them are antibodies against the specific subunits as well as antibodies recognizing common antigenic determinants. These have all been characterized according to the following properties: ELISA (enzyme‐linked immunosorbant assay) testing against each subunit, immunoblots against enriched neurofilament preparation, immunoblots of cyanogen bromide or chymotrypsin‐treated neurofilaments, immunofluorescence with PC12 cells, and immunohistochemistry of cerebellum. Whereas the antibodies against the NF68 and NF150 appear to react with single cyanogen bromide fragments, the antibodies against the NF200 react with multiple cyanogen bromide fragments. These data are consistent with the hypothesis that the NF200 is partially computed of several repeated structural determinants. Furthermore, all of the antibodies that react with the NF200 recognize the solubilized “sidearm” domain from limited chymotryptic digestions. The locations of the common and variable domains of the three subunits are discussed in light of these results.
KW - Monoclonal antibodies
KW - intermediate filaments
KW - neurofilaments
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U2 - 10.1002/jcb.240270212
DO - 10.1002/jcb.240270212
M3 - Article
C2 - 2580851
AN - SCOPUS:0021918232
SN - 0730-2312
VL - 27
SP - 181
EP - 187
JO - Journal of Cellular Biochemistry
JF - Journal of Cellular Biochemistry
IS - 2
ER -