Revision 5
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

IF-IC, FC-FP

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

Source/Isotype:

Rabbit IgG

UniProt ID:

#P04150

Entrez-Gene Id:

2908

Product Information

Product Usage Information

Application Dilution
Immunofluorescence (Immunocytochemistry) 1:400 - 1:800
Flow Cytometry (Fixed/Permeabilized) 1:50

Storage

Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.

Specificity / Sensitivity

Glucocorticoid Receptor (D8H2) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) recognizes endogenous levels of total glucocorticoid receptor protein. Based upon sequence alignment, this antibody is predicted to cross-react with all known alternative translation start site generated isoforms of glucocorticoid receptor-α and glucocorticoid receptor-β. This antibody does not cross-react with mineralocorticoid receptor.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu378 of human glucocorticoid receptor protein.

Product Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 488 fluorescent dye and tested in-house for direct immunofluorescent analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated Glucocorticoid Receptor (D8H2) XP® Rabbit mAb #3660.

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).

  1. Yamamoto, K.R. (1985) Annu. Rev. Genet 19, 209-52.
  2. Necela, B.M. and Cidlowski, J.A. (2003) Trends Pharmacol. Sci. 24, 58-61.
  3. Wang, Z. et al. (2002) J. Biol. Chem. 277, 26573-80.
  4. Rogatsky, I. et al. (1998) J. Biol. Chem. 273, 14315-21.
  5. Krstic, M. D. et al. (1997) Mol. Cell. Biol. 17, 3947-54.

Species Reactivity

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

Applications Key

IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
This product is provided under an intellectual property license from Life Technologies Corporation. The transfer of this product is conditioned on the buyer using the purchased product solely in research conducted by the buyer, excluding contract research or any fee for service research, and the buyer must not (1) use this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; or (c) manufacturing or quality assurance or quality control, and/or (2) sell or transfer this product or its components for resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or [email protected].
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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Revision 5
#12007

Glucocorticoid Receptor (D8H2) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate)

Immunofluorescence Image 1: Glucocorticoid Receptor (D8H2) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) Expand Image
使用 Glucocorticoid Receptor (D8H2) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate)(绿色),对在含 5% 活性炭剥离的 FBS 的无酚红培养基中生长 2 天,随后未经处理(左图)或用 地塞米松(100 nM,2 小时;右图)处理的 HeLa 细胞进行共聚焦免疫荧光分析。使用 DY-554 phalloidin(红色)标记肌动蛋白纤丝。
Flow Cytometry Image 1: Glucocorticoid Receptor (D8H2) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) Expand Image
人全血细胞按照 Cell Signaling Technology 的流式细胞术(备选)实验步骤进行固定、裂解和通透处理,并用 CD3-PE、CD19-APC 和 Glucocorticoid Receptor (D8H2) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) 染色。对描述糖皮质激素平均荧光强度的柱状图使用 CD3(蓝色)和 CD19(绿色)细胞群门控,并以 Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 488 Conjugate) #2975(红色)作为对照。