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
|
Holdings |
Library |
Call Number |
Additional Info |
Location |
Last Modified |
Checkout Status |
NTIS |
PB92-209345 |
Some EPA libraries have a fiche copy filed under the call number shown. |
|
07/26/2022 |
|
Collation |
8p |
Abstract |
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. |
Supplementary Notes |
Pub. in Analytical Biochemistry, v196 n2 p296-301, 1 Aug 91. Sponsored by Health Effects Research Lab., Research Triangle Park, NC. |
NTIS Title Notes |
Journal article. |
Title Annotations |
Reprint: 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. |
Category Codes |
57F; 57B |
NTIS Prices |
PC A02/MF A01 |
Primary Description |
600/10 |
Document Type |
NT |
Cataloging Source |
NTIS/MT |
Control Number |
226925138 |
Origin |
NTIS |
Type |
CAT |