Record Display for the EPA National Library Catalog


Main Title Quantitative Dot-Immunobinding Assay for Proteins Using Nitrocellulose Membrane Filters.
Author Jahn, R. ; Schiebler, W. ; Greengard, P. ;
CORP Author Rockefeller Univ., New York.;Health Effects Research Lab., Research Triangle Park, NC.
Year Published 1984
Report Number EPA-R-810608; EPA/600/J-84/382;
Stock Number PB87-146205
Additional Subjects Meetings ; Protein ; Quantitative analysis ; Rats ; Brain ; Immunoassay
Library Call Number Additional Info Location Last
NTIS  PB87-146205 Some EPA libraries have a fiche copy filed under the call number shown. 07/26/2022
Collation 7p
An immunoassay method is described for the quantitative determination of synapsin I (protein I) and of a 36,000-dalton membrane protein from rat brain synaptic vesicles. The samples are spotted on nitrocellulose membrane filters, incubated sequentially with specific antibodies and -125 I-labeled protein A, and assayed for radioactivity in a gamma scintillation counter. Conditions have been established to prevent losses of protein from the sheets during processing, to quench background radioactivity, and to adjust the sensitivity to the range desired. A large number of samples can be handled in parallel. The assay does not require iodination of the antigen and is accurate even with crude tissue samples. Standard curves were linear over a 20- to 50-fold range. The sensitivity of the method is such that 10 pmol of synapsin I and 50 ng of total vesicle membrane protein could be measured with accuracy. The method should prove useful for a wide range of proteins.