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Main Title Measurement of Tyrosine Hydroxylase Apoenzyme Protein by Enzyme-Linked Immunosorbent Assay (ELISA): Effects of 1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine (MPTP) on Striatal Tyrosine Hydroxylase Activity and Content.
Author Reinhard, J. F. ; O'Callaghan, J. P. ;
CORP Author Wellcome Research Labs., Research Triangle Park, NC.;Health Effects Research Lab., Research Triangle Park, NC.
Publisher c1991
Year Published 1991
Report Number EPA/600/J-92/272;
Stock Number PB92-209345
Additional Subjects Tyrosine hydroxylase ; Enzyme-linked immunosorbent assay ; Corpus striatum ; Monoclonal antibodies ; Mice ; IgG ; Reagents ; Immunoenzyme techniques ; Horseradish peroxidase ; Antibody specificity ; Reprints ; Methylphenyl tetrahydropyridine
Library Call Number Additional Info Location Last
NTIS  PB92-209345 Some EPA libraries have a fiche copy filed under the call number shown. 07/26/2022
Collation 8p
An enzyme-linked immunosorbent assay (ELISA) has been developed for tyrosine hydroxylase (TH). The method uses a polyclonal antibody to trap TH, a monoclonal antibody to bind the immobilized TH, a biotinylated, anti-mouse immunoglobulin to bind the monoclonal antibody, and streptavidin covalently coupled to horseradish peroxidase (SA-HRP). The incubations are performed at 37 C and the antigen-antibody complex detected colorometrically following incubation with an HRP substrate. The method detects less than 1 ng (16 fmol) of TH and can be performed in 3 hours or less on multiples of 96. The high specificity of the assay is attributed to the use of both polyclonal and monoclonal antibodies, each of which are specific for TH. Data acquisition and reduction is rapid (10 seconds/plate) and linked directly to a common desktop computer. Levels of TH protein average 1 ng/micrograms protein in striatum and, following treatment with the neurotoxicant MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine), are decreased to a similar extent as is catalytic activity. In contrast, MPTP did not alter TH homospecific activity. The monoamine oxidase B inhibitor deprenyl blocked both the decrease in activity and the decrease in immunoreactive protein caused by MPTP.