Immunological study of a neurofilament protein variant (S150) with polyclonal and monoclonal antibodies.
Ten polyclonal neurofilament antibodies were tested for domain specificity with immunoblots of chymotrypsin digests of a neurofilament protein of 150 kDa (NF 150K). In contrast to most monoclonal antibodies previously reported, the five polyclonal antibodies which showed domain specificity reacted with the 40 kDa alpha-helical rod domain of the molecule. (With one exception, monoclonal antibodies reacted with the 100 kDa carboxy-terminal peripheral domain). Of these ten polyclonal antibodies only two reacted with an isoelectric variant of NK 150K (S150) isolated by Liem and collaborators (Wong, J., Hutchison, S.B. and Liem, R.K.H. (1984) J. Biol. Chem. 259, 10867-10874) from bovine brain. 13 monoclonal antibodies were also tested for reactivity with S150 protein. With one exception, none of these antibodies reacted with this variant, not even a monoclonal antibody which we have previously shown to react with a non-phosphorylated epitope located in the rod domain of NF 150K. We suggest that either there are modifications other than dephosphorylation in the S150 isoelectric variant or, alternatively, that it is not derived from NF 150K.
D000818:Animals; D000911:Antibodies, Monoclonal; D001923:Brain Chemistry; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000032:analysis; D007124:Immunoenzyme Techniques; D007381:Intermediate Filament Proteins / Q000032:analysis*; D016900:Neurofilament Proteins
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007381:Intermediate Filament Proteins; D016900:Neurofilament Proteins