Multi-well plate immunoassays for measuring signaling protein activations/deactivations and membrane vs. intracellular receptor levels

We developed fixed-cell multi-well plate immunoassays that increase the throughput and ease of quantification for questions formerly assessed by immunoblot scanning. The assays make use of the now abundant antibodies designed to recognize receptor subtypes and posttranslationally modified signaling...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 1204; p. 123
Main Authors Watson, Cheryl S, Jeng, Yow-Jiun, Bulayeva, Nataliya N, Finnerty, Celeste C, Koong, Luke Y, Zivadinovic, Dragoslaca, Alyea, Rebecca A, Midoro-Horiuti, Terumi, Goldblum, Randall M, Anastasio, Noelle C, Cunningham, Kathryn A, Seitz, Patricia K, Smith, Thressa D
Format Journal Article
LanguageEnglish
Published United States 01.01.2014
Subjects
Animals
Cell Line
Cells, Cultured
Enzyme Activation
Enzyme Assays - methods
Humans
Immunoassay - methods
MAP Kinase Signaling System
Receptors, Estrogen - analysis
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Summary:We developed fixed-cell multi-well plate immunoassays that increase the throughput and ease of quantification for questions formerly assessed by immunoblot scanning. The assays make use of the now abundant antibodies designed to recognize receptor subtypes and posttranslationally modified signaling proteins. By optimizing permeabilization and fixation conditions, mainly based on specific cell types, the assay can be adapted to the study of many different antigens of importance to hormonal and neurotransmitter signaling scenarios.
ISSN:1940-6029
DOI:10.1007/978-1-4939-1346-6_11