Product Pathways - Nuclear Receptor Signaling
Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb #12041
|12041S||100 µl (10 western blots)||---||In Stock||---|
|12041P||40 µl (4 western blots)||---||In Stock||---|
|12041||carrier free and custom formulation / quantity||email request|
Already purchased this product? Write a Review.
|W||1:1000||Human, Mouse, Rat, Monkey||Endogenous||94, 91||Rabbit IgG|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry, ChIP=Chromatin IP
* Product-specific protocol.
Specificity / Sensitivity
Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb recognizes endogenous levels of total GR protein. This antibody reacts with GR-α and GR-β but does not cross-react with mineralocorticoid receptor.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a recombinant protein specific to the amino terminus of human GR protein.
Western blot analysis of extracts from various cell lines using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb.
Western blot analysis of extracts from 293T cells, either mock transfected (-) or transfected with constructs expressing Myc/DDK-tagged full-length human glucocorticoid receptor-α (hGRα-Myc/DDK; +) or Myc/DDK-tagged full-length human mineralocorticoid receptor (hMR-Myc/DDK; +), using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb (upper) or DYKDDDDK Tag Antibody #2368 (lower).
Immunoprecipitation of glucocorticoid receptor from HeLa cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse stomach using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded HeLa cell pellets, untreated (left) or dexamethasone-treated (right), using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb.
Human whole blood was fixed, lysed, and permeabilized as per the Cell Signaling Technology Flow Cytometry (Alternate) Protocol, and stained with CD3-PE, CD19-APC, and Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb. B cell (green) and T cell (blue) population gates (left) were applied to a histogram depicting the mean fluorescence intensity of glucocorticoid receptor, compared to a nonspecific negative control antibody (red; right). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Confocal immunofluorescent analysis of HeLa cells, grown in phenol red-free media containing 5% charcoal-stripped FBS for 2 d and either untreated (left) or treated with dexamethasone (100 nM, 2 hr; right), using Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
A549 cells were cultured in media with 5% charcoal-stripped FBS for 3 d and then either untreated (left panel) or dexamethasone-treated (100 nM, 1 hr; right panel). Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 cells and 10 µl of Glucocorticoid Receptor (D6H2L) XP® Rabbit mAb or 2 µl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human SLC19A2 Promoter Primers #7681, human MT2A promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
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). Indeed Ser211 of human GR is phosphorylated to a greater extent in the presence of hormone, and biochemical fractionation studies following hormone treatment indicate that Ser211-phosphorylated GR is found in the nucleus (3). Thus, Ser211 phosphorylation is a biomarker for activated GR in vivo. An added layer of complexity to GR signaling lies in the ability of multiple isoforms to be generated through both alternative splicing and the use of alternative translation intiation start sites, thus increasing the repertoire of functional signaling homo- and heterodimers (6,7).
- Yamamoto, K.R. (1985) Annu. Rev. Genet 19, 209-252.
- Necela, B.M. and Cidlowski, J.A. (2003) Trends Pharmacol. Sci. 24, 58-61.
- Wang, Z. et al. (2002) J. Biol. Chem. 277, 26573-26580.
- Rogatsky, I. et al. (1998) J. Biol. Chem. 273, 14315-14321.
- Krstic, M. D. et al. (1997) Mol. Cell. Biol. 17, 3947-3954.
- Yudt, M.R. and Cidlowski, J.A. (2001) Mol Endocrinol 15, 1093-103.
- Lu, N.Z. and Cidlowski, J.A. (2005) Mol Cell 18, 331-42.
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
- 4161 Phospho-Glucocorticoid Receptor (Ser211) Antibody
- 5153 Androgen Receptor (D6F11) XP® Rabbit mAb
- 8757 Progesterone Receptor A/B (D8Q2J) XP® Rabbit mAb
- 3660 Glucocorticoid Receptor (D8H2) XP® Rabbit mAb
- 3157 Progesterone Receptor B (C1A2) Rabbit mAb
- 9002 SimpleChIP® Enzymatic Chromatin IP Kit (Agarose Beads)
- 9003 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads)
- 9004 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Agarose Beads)
- 9005 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads)
- 9006 ChIP-Grade Protein G Magnetic Beads
- 9007 ChIP-Grade Protein G Agarose Beads
- 7681 SimpleChIP® Human SLC19A2 Promoter Primers
- 4486 SimpleChIP® Human α Satellite Repeat Primers
- 7017 6-Tube Magnetic Separation Rack
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 9999 Nonfat Dry Milk
- 3900 Rabbit (DA1E) mAb IgG XP® Isotype Control
- 2729 Normal Rabbit IgG
- 9863 Protein A Agarose Beads
- 6508 SignalSilence® Glucocorticoid Receptor siRNA I
- 7699 SignalSilence® Glucocorticoid Receptor siRNA II
- 8112 SignalStain® Antibody Diluent
- 8114 SignalStain® Boost IHC Detection Reagent (HRP, Rabbit)
- 5425 Normal Goat Serum
- 9997 Tris Buffered Saline with Tween® 20 (TBST-10X)
- 8059 SignalStain® DAB Substrate Kit
For Research Use Only. Not For Use In Diagnostic Procedures.
XP® is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor® is a registered trademark of Molecular Probes, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.