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

Cheryl S. Watson; Yow Jiun Jeng; Nataliya N. Bulayeva; Celeste C. Finnerty; Luke Y. Koong; Dragoslaca Zivadinovic; Rebecca A. Alyea; Terumi Midoro-Horiuti; Randall M. Goldblum; Noelle C. Anastasio; Kathryn A. Cunningham; Patricia K. Seitz; Thressa D. Smith

doi:10.1007/978-1-4939-1346-6_11

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

Cheryl S. Watson, Yow Jiun Jeng, Nataliya N. Bulayeva, Celeste C. Finnerty, Luke Y. Koong, Dragoslaca Zivadinovic, Rebecca A. Alyea, Terumi Midoro-Horiuti, Randall M. Goldblum, Noelle C. Anastasio, Kathryn A. Cunningham, Patricia K. Seitz, Thressa D. Smith

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

Abstract

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 s ignaling scenarios.

Original language	English (US)
Pages (from-to)	123-133
Number of pages	11
Journal	Methods in Molecular Biology
Volume	1204
DOIs	https://doi.org/10.1007/978-1-4939-1346-6_11
State	Published - 2014
Externally published	Yes

Keywords

Cells
Immunoblots
Posttranslational modifications
Quantification
Throughput
Tissues

ASJC Scopus subject areas

Molecular Biology
Genetics

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10.1007/978-1-4939-1346-6_11

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Watson, C. S., Jeng, Y. J., Bulayeva, N. N., Finnerty, C. C., Koong, L. Y., Zivadinovic, D., Alyea, R. A., Midoro-Horiuti, T., Goldblum, R. M., Anastasio, N. C., Cunningham, K. A., Seitz, P. K., & Smith, T. D. (2014). Multi-well plate immunoassays for measuring signaling protein activations/deactivations and membrane vs. intracellular receptor levels. Methods in Molecular Biology, 1204, 123-133. https://doi.org/10.1007/978-1-4939-1346-6_11

Watson, CS, Jeng, YJ, Bulayeva, NN, Finnerty, CC, Koong, LY, Zivadinovic, D, Alyea, RA, Midoro-Horiuti, T, Goldblum, RM, Anastasio, NC , Cunningham, KA, Seitz, PK & Smith, TD 2014, 'Multi-well plate immunoassays for measuring signaling protein activations/deactivations and membrane vs. intracellular receptor levels', Methods in Molecular Biology, vol. 1204, pp. 123-133. https://doi.org/10.1007/978-1-4939-1346-6_11

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abstract = "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 s ignaling scenarios.",

keywords = "Cells, Immunoblots, Posttranslational modifications, Quantification, Throughput, Tissues",

author = "Watson, {Cheryl S.} and Jeng, {Yow Jiun} and Bulayeva, {Nataliya N.} and Finnerty, {Celeste C.} and Koong, {Luke Y.} and Dragoslaca Zivadinovic and Alyea, {Rebecca A.} and Terumi Midoro-Horiuti and Goldblum, {Randall M.} and Anastasio, {Noelle C.} and Cunningham, {Kathryn A.} and Seitz, {Patricia K.} and Smith, {Thressa D.}",

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year = "2014",

doi = "10.1007/978-1-4939-1346-6_11",

language = "English (US)",

volume = "1204",

pages = "123--133",

journal = "Methods in Molecular Biology",

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AU - Watson, Cheryl S.

AU - Jeng, Yow Jiun

AU - Bulayeva, Nataliya N.

AU - Finnerty, Celeste C.

AU - Koong, Luke Y.

AU - Zivadinovic, Dragoslaca

AU - Alyea, Rebecca A.

AU - Midoro-Horiuti, Terumi

AU - Goldblum, Randall M.

AU - Anastasio, Noelle C.

AU - Cunningham, Kathryn A.

AU - Seitz, Patricia K.

AU - Smith, Thressa D.

N1 - Publisher Copyright: © Springer Science+Business Media New York 2014.

PY - 2014

Y1 - 2014

N2 - 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 s ignaling scenarios.

AB - 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 s ignaling scenarios.

KW - Cells

KW - Immunoblots

KW - Posttranslational modifications

KW - Quantification

KW - Throughput

KW - Tissues

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

Abstract

Keywords

ASJC Scopus subject areas

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