A quantitative dot-immunobinding assay for proteins using nitrocellulose membrane filters.
- 1 March 1984
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 81 (6) , 1684-1687
- https://doi.org/10.1073/pnas.81.6.1684
Abstract
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 125I-labeled protein A and assayed for radioactivity in a gamma scintillation counter. Conditions were 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 20-50-fold range. 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.This publication has 18 references indexed in Scilit:
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