Binding of Bodian's silver and monoclonal antibodies to defined regions of human neurofilament subunits: Bodian's silver reacts with a highly charged unique domain of neurofilaments.
Cleavage at cysteine and chymotrypsin digestion were applied to two human neurofilament (NF) subunits, low- and high-molecular-weight NF (NF-L and NF-H), to locate the regions reacting with Bodian's silver stain and with several monoclonal antibodies, including NF-specific antibodies and one that recognizes all intermediate filaments (anti-IFA). Our findings indicate that whereas anti-IFA recognizes the highly conserved rod domain, all the NF-specific antibodies, as well as Bodian's silver, react with the carboxy-terminal tailpiece of NF subunits. The silver binding sites in NF-L are located in a carboxy-terminal 12-Kd chymotrypsin fragment, a highly charged, unique domain of NF.
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Autilio-Gambetti
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Crane
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Gambetti
D000911:Antibodies, Monoclonal; D001665:Binding Sites; D002918:Chymotrypsin / Q000378:metabolism; D003599:Cytoskeleton / Q000648:ultrastructure*; D004591:Electrophoresis, Polyacrylamide Gel; D006651:Histocytochemistry; D006801:Humans; D007381:Intermediate Filament Proteins / Q000032:analysis*; D007382:Intermediate Filaments / Q000648:ultrastructure*; D046911:Macromolecular Substances; D008722:Methods; D008970:Molecular Weight; D016900:Neurofilament Proteins; D010446:Peptide Fragments / Q000378:metabolism; D012834:Silver / Q000378:metabolism*; D013194:Staining and Labeling
D000911:Antibodies, Monoclonal; D007381:Intermediate Filament Proteins; D046911:Macromolecular Substances; D016900:Neurofilament Proteins; D010446:Peptide Fragments; D012834:Silver; D002918:Chymotrypsin