文章基本信息

标题：A quantitative dot-immunobinding assay for proteins using nitrocellulose membrane filters
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作者：R Jahn ; W Schiebler ; P Greengard 等
期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：1984
卷号：81
期号：6
页码：1684-1687
DOI：10.1073/pnas.81.6.1684
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：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 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.