title: Multi-well plate immunoassays for measuring signaling protein activations/deactivations and membrane vs. intracellular receptor levels.
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 signaling scenarios.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, 0645 312 Basic Science Building, Galveston, TX, 77555, USA, cswatson@utmb.edu.
authors.forename: Cheryl S
authors.initials: CS
authors.lastname: Watson
authors.forename: Yow-Jiun
authors.initials: YJ
authors.lastname: Jeng
authors.forename: Nataliya N
authors.initials: NN
authors.lastname: Bulayeva
authors.forename: Celeste C
authors.initials: CC
authors.lastname: Finnerty
authors.forename: Luke Y
authors.initials: LY
authors.lastname: Koong
authors.forename: Dragoslaca
authors.initials: D
authors.lastname: Zivadinovic
authors.forename: Rebecca A
authors.initials: RA
authors.lastname: Alyea
authors.forename: Terumi
authors.initials: T
authors.lastname: Midoro-Horiuti
authors.forename: Randall M
authors.initials: RM
authors.lastname: Goldblum
authors.forename: Noelle C
authors.initials: NC
authors.lastname: Anastasio
authors.forename: Kathryn A
authors.initials: KA
authors.lastname: Cunningham
authors.forename: Patricia K
authors.initials: PK
authors.lastname: Seitz
authors.forename: Thressa D
authors.initials: TD
authors.lastname: Smith
mesh_terms: D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D004789:Enzyme Activation; D057075:Enzyme Assays / Q000379:methods; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D020935:MAP Kinase Signaling System; D011960:Receptors, Estrogen / Q000032:analysis*
chemical_list: D011960:Receptors, Estrogen