Revision 2

#25665

Store at -20C

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
SCA-ITS3

Reactivity:
H M

Sensitivity:
Endogenous

Source/Isotype:
Rabbit IgG

UniProt ID:
#P04150

Entrez-Gene Id:
2908

Storage

Supplied in PBS (pH 7.2), 2 mM EDTA, 0.05% Triton X-100, 2 mg/mL BSA, and 50% glycerol. Store at -20°C. Do not aliquot the antibody.

Specificity/Sensitivity

Phospho-Glucocorticoid Receptor (Ser226) (D9D3V) Rabbit Monoclonal Antibody (InTraSeq™ 3' Conjugate 3036) recognizes endogenous levels of glucocorticoid receptor protein only when phosphorylated at Ser226.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser226 of human glucocorticoid receptor protein.

Background

Glucocorticoid hormones control cellular proliferation, inflammation, and metabolism through their association with the glucocorticoid receptor (GR)/NR3C1, a member of the nuclear hormone receptor superfamily of transcription factors (1). GR is composed of several conserved structural elements, including a carboxy-terminal ligand-binding domain (which also contains residues critical for receptor dimerization and hormone-dependent gene transactivation), a neighboring hinge region containing nuclear localization signals, a central zinc-finger-containing DNA-binding domain, and an amino-terminal variable region that participates in ligand-independent gene transcription. In the absence of hormone, a significant population of GR is localized to the cytoplasm in an inactive form via its association with regulatory chaperone proteins, such as HSP90, HSP70, and FKBP52. On hormone binding, GR is released from the chaperone complex and translocates to the nucleus as a dimer to associate with specific DNA sequences termed glucocorticoid response elements (GREs), thereby enhancing or repressing transcription of specific target genes (2). It was demonstrated that GR-mediated transcriptional activation is modulated by phosphorylation (3-5). Although GR can be basally phosphorylated in the absence of hormone, it becomes hyperphosphorylated upon binding receptor agonists. It has been suggested that hormone-dependent phosphorylation of GR may determine target promoter specificity, cofactor interaction, strength and duration of receptor signaling, receptor stability, and receptor subcellular localization (3).

Phosphorylation of GR at serine 226 by JNK enhances nuclear export after ligand depletion (6,7). Phosphorylation of various serine residues, including serine 226, also affects GR binding to different target genes, contributing to an additional layer of transcriptional regulation (8). Serine 226 phosphorylation has also been linked to depression disorders as well as inflammation (9-11).

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Applications Key

SCA-ITS3: Single Cell Analysis - InTraSeq™ 3'

Cross-Reactivity Key

H: Human M: Mouse

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

10x Genomics, 10x, Feature Barcode, and Chromium are the trademarks or registered trademarks of 10x Genomics, Inc.

InTraSeq is a trademark of Cell Signaling Technology, Inc.

Subject to patents licensed from 10x Genomics, Inc. for use with single-cell (i.e., Chromium) 10x products.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

Cell Signaling Technology Logo
Cultured human peripheral blood mononuclear cells (PBMCs) treated with Cell Stimulation Cocktail (without Protein Transport Inhibitors) (500X) #29255 (30 min, Stimulated; left panels) or treated with Cell Stimulation Cocktail (without Protein Transport Inhibitors) (500X) #29255 (30 min) then subsequently treated with Lambda Protein Phosphatase (40 min, 30°C, Stimulated + Phosphatase; right panels) were processed with the InTraSeq™ 3’ protocol. The T cell population was then selected and analyzed. The FeaturePlots in the top figures display the Phospho-Glucocorticoid Receptor (Ser226) (D9D3V) Rabbit mAb (InTraSeq™ 3' Conjugate 3036) expression on the UMAP. The FeaturePlots in the bottom figures display the Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb (InTraSeq™ 3' Conjugate 3035) #25504 expression on the UMAP. To generate similar plots, use the FeaturePlot command template from the Seurat computational package: FeaturePlot(your_object, features = your_protein, max.cutoff = "q95", min.cutoff = "q5", order = TRUE) + scale_color_gradientn(colours = c("#22578B", "#73A9D2", "grey90", "#FDB393", "#b22222"))
SCA Image 1: Phospho-Glucocorticoid Receptor (Ser226) (D9D3V) Rabbit Monoclonal Antibody (InTraSeq<sup>™</sup> 3' Conjugate 3036)
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.