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 |
|---|---|
| Pages (from-to) | 123-133 |
| Number of pages | 11 |
| Journal | Methods in molecular biology (Clifton, N.J.) |
| Volume | 1204 |
| DOIs | |
| State | Published - 2014 |
| Externally published | Yes |
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Keywords
- Cells
- Immunoblots
- Posttranslational modifications
- Quantification
- Throughput
- Tissues
ASJC Scopus subject areas
- Molecular Biology
- Genetics
Cite this
Multi-well plate immunoassays for measuring signaling protein activations/deactivations and membrane vs. intracellular receptor levels. / Watson, Cheryl S.; Jeng, Yow Jiun; Bulayeva, Nataliya N.; Finnerty, Celeste; Koong, Luke Y.; Zivadinovic, Dragoslaca; Alyea, Rebecca A.; Midoro-Horiuti, Terumi; Goldblum, Randall M.; Anastasio, Noelle; Cunningham, Kathryn; Seitz, Patricia K.; Smith, Thressa D.
In: Methods in molecular biology (Clifton, N.J.), Vol. 1204, 2014, p. 123-133.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Multi-well plate immunoassays for measuring signaling protein activations/deactivations and membrane vs. intracellular receptor levels
AU - Watson, Cheryl S.
AU - Jeng, Yow Jiun
AU - Bulayeva, Nataliya N.
AU - Finnerty, Celeste
AU - Koong, Luke Y.
AU - Zivadinovic, Dragoslaca
AU - Alyea, Rebecca A.
AU - Midoro-Horiuti, Terumi
AU - Goldblum, Randall M.
AU - Anastasio, Noelle
AU - Cunningham, Kathryn
AU - Seitz, Patricia K.
AU - Smith, Thressa D.
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
UR - http://www.scopus.com/inward/record.url?scp=84921806485&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84921806485&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-1346-6_11
DO - 10.1007/978-1-4939-1346-6_11
M3 - Article
C2 - 25182766
AN - SCOPUS:84921806485
VL - 1204
SP - 123
EP - 133
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
SN - 1064-3745
ER -