Product: Phospho-NR3C1 (Ser226) Antibody
Catalog: AF3066
Description: Rabbit polyclonal antibody to Phospho-NR3C1 (Ser226)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog
Mol.Wt.: 86kDa; 86kD(Calculated).
Uniprot: P04150
RRID: AB_2834430

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(89%), Bovine(100%), Horse(90%), Sheep(100%), Rabbit(100%), Dog(100%)
Clonality:
Polyclonal
Specificity:
Phospho-NR3C1 (Ser226) Antibody detects endogenous levels of NR3C1 only when phosphorylated at Serine 226.
RRID:
AB_2834430
Cite Format: Affinity Biosciences Cat# AF3066, RRID:AB_2834430.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

GCCR; GCR; GCR_HUMAN; glucocorticoid nuclear receptor variant 1; Glucocorticoid receptor; Glucocorticoid receptor beta isoform; GR; GRL; nr3c1; Nuclear receptor subfamily 3 group C member 1;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P04150 GCR_HUMAN:

Widely expressed including bone, stomach, lung, liver, colon, breast, ovary, pancreas and kidney (PubMed:25847991). In the heart, detected in left and right atria, left and right ventricles, aorta, apex, intraventricular septum, and atrioventricular node as well as whole adult and fetal heart (PubMed:10902803). Isoform Beta: Widely expressed including brain, bone marrow, thymus, spleen, liver, kidney, pancreas, lung, fat, skeletal muscle, heart, placenta and blood leukocytes (PubMed:7769088, PubMed:8621628). Isoform Alpha-2: Expressed at low level.

Description:
The protein encoded by this gene is a receptor for glucocorticoids and can act as both a transcription factor and a regulator of other transcription factors. The encoded protein can bind DNA as a homodimer or as a heterodimer with another protein such as the retinoid X receptor. This protein can also be found in heteromeric cytoplasmic complexes along with heat shock factors and immunophilins.
Sequence:
MDSKESLTPGREENPSSVLAQERGDVMDFYKTLRGGATVKVSASSPSLAVASQSDSKQRRLLVDFPKGSVSNAQQPDLSKAVSLSMGLYMGETETKVMGNDLGFPQQGQISLSSGETDLKLLEESIANLNRSTSVPENPKSSASTAVSAAPTEKEFPKTHSDVSSEQQHLKGQTGTNGGNVKLYTTDQSTFDILQDLEFSSGSPGKETNESPWRSDLLIDENCLLSPLAGEDDSFLLEGNSNEDCKPLILPDTKPKIKDNGDLVLSSPSNVTLPQVKTEKEDFIELCTPGVIKQEKLGTVYCQASFPGANIIGNKMSAISVHGVSTSGGQMYHYDMNTASLSQQQDQKPIFNVIPPIPVGSENWNRCQGSGDDNLTSLGTLNFPGRTVFSNGYSSPSMRPDVSSPPSSSSTATTGPPPKLCLVCSDEASGCHYGVLTCGSCKVFFKRAVEGQHNYLCAGRNDCIIDKIRRKNCPACRYRKCLQAGMNLEARKTKKKIKGIQQATTGVSQETSENPGNKTIVPATLPQLTPTLVSLLEVIEPEVLYAGYDSSVPDSTWRIMTTLNMLGGRQVIAAVKWAKAIPGFRNLHLDDQMTLLQYSWMFLMAFALGWRSYRQSSANLLCFAPDLIINEQRMTLPCMYDQCKHMLYVSSELHRLQVSYEEYLCMKTLLLLSSVPKDGLKSQELFDEIRMTYIKELGKAIVKREGNSSQNWQRFYQLTKLLDSMHEVVENLLNYCFQTFLDKTMSIEFPEMLAEIITNQIPKYSNGNIKKLLFHQK

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Bovine
100
Sheep
100
Dog
100
Rabbit
100
Horse
90
Pig
89
Xenopus
0
Zebrafish
0
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P04150 As Substrate

Site PTM Type Enzyme
S6 Phosphorylation
T8 Phosphorylation
Y30 Phosphorylation
T32 Phosphorylation
R34 Methylation
T38 Phosphorylation
S44 Phosphorylation
S45 Phosphorylation
S54 Phosphorylation
S56 Phosphorylation
K57 Sumoylation
K57 Ubiquitination
K67 Sumoylation
K67 Ubiquitination
S79 Phosphorylation
Y89 Phosphorylation
T93 Phosphorylation
T95 Phosphorylation
K96 Sumoylation
S113 Phosphorylation
S125 Phosphorylation
S132 Phosphorylation
T133 Phosphorylation
S134 Phosphorylation P31749 (AKT1)
K140 Ubiquitination
S141 Phosphorylation
K154 Acetylation
K154 Ubiquitination
S164 Phosphorylation
K171 Ubiquitination
S203 Phosphorylation Q00535 (CDK5) , Q16539 (MAPK14)
K206 Ubiquitination
T208 Phosphorylation
S211 Phosphorylation Q16539 (MAPK14) , Q00535 (CDK5)
S226 Phosphorylation Q16539 (MAPK14) , P45983 (MAPK8) , Q00535 (CDK5) , P28482 (MAPK1) , Q15759 (MAPK11) , P27361 (MAPK3) , P53779 (MAPK10) , P45984 (MAPK9)
S234 Phosphorylation
S241 Phosphorylation
K254 Sumoylation
S267 Phosphorylation
T272 Phosphorylation
K277 Sumoylation
K277 Ubiquitination
K280 Ubiquitination
T288 Phosphorylation
K293 Sumoylation
K293 Ubiquitination
K296 Ubiquitination
Y301 Phosphorylation
S305 Phosphorylation
S317 Phosphorylation
K348 Sumoylation
S390 Phosphorylation
S404 Phosphorylation P49841 (GSK3B)
K480 Acetylation
K492 Acetylation
K494 Acetylation
K495 Acetylation
K498 Sumoylation
K498 Ubiquitination
S508 Phosphorylation
K677 Ubiquitination
K681 Sumoylation
K681 Ubiquitination
K695 Ubiquitination
K699 Ubiquitination
K703 Sumoylation
K770 Ubiquitination
K771 Ubiquitination

Research Backgrounds

Function:

Receptor for glucocorticoids (GC). Has a dual mode of action: as a transcription factor that binds to glucocorticoid response elements (GRE), both for nuclear and mitochondrial DNA, and as a modulator of other transcription factors. Affects inflammatory responses, cellular proliferation and differentiation in target tissues. Involved in chromatin remodeling. Plays a role in rapid mRNA degradation by binding to the 5' UTR of target mRNAs and interacting with PNRC2 in a ligand-dependent manner which recruits the RNA helicase UPF1 and the mRNA-decapping enzyme DCP1A, leading to RNA decay. Could act as a coactivator for STAT5-dependent transcription upon growth hormone (GH) stimulation and could reveal an essential role of hepatic GR in the control of body growth (By similarity).

Has transcriptional activation and repression activity. Mediates glucocorticoid-induced apoptosis. Promotes accurate chromosome segregation during mitosis. May act as a tumor suppressor. May play a negative role in adipogenesis through the regulation of lipolytic and antilipogenic gene expression (By similarity).

Acts as a dominant negative inhibitor of isoform Alpha. Has intrinsic transcriptional activity independent of isoform Alpha when both isoforms are coexpressed. Loses this transcription modulator function on its own. Has no hormone-binding activity. May play a role in controlling glucose metabolism by maintaining insulin sensitivity (By similarity). Reduces hepatic gluconeogenesis through down-regulation of PEPCK in an isoform Alpha-dependent manner. Directly regulates STAT1 expression in isoform Alpha-independent manner.

Has lower transcriptional activation activity than isoform Alpha. Exerts a dominant negative effect on isoform Alpha trans-repression mechanism.

Increases activity of isoform Alpha.

More effective than isoform Alpha in transcriptional activation, but not repression activity.

Has transcriptional activation activity.

Has transcriptional activation activity.

Has transcriptional activation activity.

Has highest transcriptional activation activity of all isoforms created by alternative initiation. Has transcriptional repression activity. Mediates glucocorticoid-induced apoptosis.

Has transcriptional activation activity.

Has transcriptional activation activity.

Has lowest transcriptional activation activity of all isoforms created by alternative initiation. Has transcriptional repression activity.

PTMs:

Acetylation by CLOCK reduces its binding to glucocorticoid response elements and its transcriptional activity.

Increased proteasome-mediated degradation in response to glucocorticoids. Isoform Alpha-B appears to be more susceptible to proteolytic degradation than isoform Alpha.

Phosphorylated in the absence of hormone; becomes hyperphosphorylated in the presence of glucocorticoid. The Ser-203, Ser-226 and Ser-404-phosphorylated forms are mainly cytoplasmic, and the Ser-211-phosphorylated form is nuclear. Phosphorylation at Ser-211 increases transcriptional activity. Phosphorylation at Ser-203, Ser-226 and Ser-404 decreases signaling capacity. Phosphorylation at Ser-404 may protect from glucocorticoid-induced apoptosis. Phosphorylation at Ser-203 and Ser-211 is not required in regulation of chromosome segregation. May be dephosphorylated by PPP5C, attenuates NR3C1 action (By similarity).

Sumoylation at Lys-277 and Lys-293 negatively regulates its transcriptional activity. Sumoylation at Lys-703 positively regulates its transcriptional activity in the presence of RWDD3 (By similarity). Sumoylation at Lys-277 and Lys-293 is dispensable whereas sumoylation at Lys-703 is critical for the stimulatory effect of RWDD3 on its transcriptional activity (By similarity). Heat shock increases sumoylation in a RWDD3-dependent manner (By similarity).

Ubiquitinated; restricts glucocorticoid-mediated transcriptional signaling.

Subcellular Location:

Cytoplasm. Nucleus. Mitochondrion. Cytoplasm>Cytoskeleton>Spindle. Cytoplasm>Cytoskeleton>Microtubule organizing center>Centrosome.
Note: After ligand activation, translocates from the cytoplasm to the nucleus. In the presence of NR1D1 shows a time-dependent subcellular localization, localizing to the cytoplasm at ZT8 and to the nucleus at ZT20 (By similarity). Lacks this diurnal pattern of localization in the absence of NR1D1, localizing to both nucleus and the cytoplasm at ZT8 and ZT20 (By similarity).

Nucleus. Cytoplasm.
Note: Expressed predominantly in the nucleus with some expression also detected in the cytoplasm.

Nucleus. Cytoplasm.
Note: After ligand activation, translocates from the cytoplasm to the nucleus.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Widely expressed including bone, stomach, lung, liver, colon, breast, ovary, pancreas and kidney. In the heart, detected in left and right atria, left and right ventricles, aorta, apex, intraventricular septum, and atrioventricular node as well as whole adult and fetal heart. Isoform Beta: Widely expressed including brain, bone marrow, thymus, spleen, liver, kidney, pancreas, lung, fat, skeletal muscle, heart, placenta and blood leukocytes. Isoform Alpha-2: Expressed at low level.

Subunit Structure:

Heteromultimeric cytoplasmic complex with HSP90AA1, HSPA1A/HSPA1B, and FKBP5 or another immunophilin such as PPID, STIP1, or the immunophilin homolog PPP5C. Upon ligand binding FKBP5 dissociates from the complex and FKBP4 takes its place, thereby linking the complex to dynein and mediating transport to the nucleus, where the complex dissociates (By similarity). Probably forms a complex composed of chaperones HSP90 and HSP70, co-chaperones CDC37, PPP5C, TSC1 and client protein TSC2, CDK4, AKT, RAF1 and NR3C1; this complex does not contain co-chaperones STIP1/HOP and PTGES3/p23. Directly interacts with UNC45A. Binds to DNA as a homodimer, and as heterodimer with NR3C2 or the retinoid X receptor. Binds STAT5A and STAT5B homodimers and heterodimers (By similarity). Interacts with NRIP1, POU2F1, POU2F2 and TRIM28 (By similarity). Interacts with several coactivator complexes, including the SMARCA4 complex, CREBBP/EP300, TADA2L (Ada complex) and p160 coactivators such as NCOA2 and NCOA6. Interaction with BAG1 inhibits transactivation. Interacts with HEXIM1, PELP1 and TGFB1I1. Interacts with NCOA1. Interacts with NCOA3, SMARCA4, SMARCC1, SMARCD1, and SMARCE1 (By similarity). Interacts with CLOCK, CRY1 and CRY2 in a ligand-dependent fashion. Interacts with CIART (By similarity). Interacts with RWDD3 (By similarity). Interacts with UBE2I/UBC9 and this interaction is enhanced in the presence of RWDD3 (By similarity). Interacts with GRIP1. Interacts with NR4A3 (via nuclear receptor DNA-binding domain), represses transcription activity of NR4A3 on the POMC promoter Nur response element (NurRE). Directly interacts with PNRC2 to attract and form a complex with UPF1 and DCP1A; the interaction leads to rapid mRNA degradation. Interacts with GSK3B. Interacts with FNIP1 and FNIP2. Interacts (via C-terminus) with HNRNPU (via C-terminus). Interacts with MCM3AP. Interacts (via domain NR LBD) with HSP90AA1 and HSP90AB1 (By similarity). In the absence of hormonal ligand, interacts with TACC1.

Family&Domains:

Composed of three domains: a modulating N-terminal domain, a DNA-binding domain and a C-terminal ligand-binding domain (PubMed:3841189). The ligand-binding domain is required for correct chromosome segregation during mitosis although ligand binding is not required (PubMed:25847991).

Belongs to the nuclear hormone receptor family. NR3 subfamily.

Research Fields

· Environmental Information Processing > Signaling molecules and interaction > Neuroactive ligand-receptor interaction.

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