Phospho-RUNX2 (Ser275) Antibody - #AF7379

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Product: Phospho-RUNX2 (Ser275) Antibody
Catalog: AF7379
Description: Rabbit polyclonal antibody to Phospho-RUNX2 (Ser275)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Chicken, Xenopus
Mol.Wt.: 50-60kd; 57kD(Calculated).
Uniprot: Q13950
RRID: AB_2843819

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 100ul $350 In stock
 200ul $450 In stock

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Related Downloads
MS Report
HPLC Report
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

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(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Chicken(100%), Xenopus(100%)
Clonality:
Polyclonal
Specificity:
Phospho-RUNX2 (Ser275) Antibody detects endogenous levels of RUNX2 only when phosphorylated at Ser275.
RRID:
AB_2843819
Cite Format: Affinity Biosciences Cat# AF7379, RRID:AB_2843819.
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

Acute myeloid leukemia 3 protein; Alpha subunit 1; AML3; CBF alpha 1; CBF-alpha-1; CBFA1; CCD; CCD1; Cleidocranial dysplasia 1; Core binding factor; Core binding factor runt domain alpha subunit 1; Core binding factor subunit alpha 1; Core-binding factor subunit alpha-1; MGC120022; MGC120023; Oncogene AML 3; Oncogene AML-3; OSF 2; OSF-2; OSF2; Osteoblast specific transcription factor 2; Osteoblast-specific transcription factor 2; OTTHUMP00000016533; PEA2 alpha A; PEA2-alpha A; PEA2aA; PEBP2 alpha A; PEBP2-alpha A; PEBP2A1; PEBP2A2; PEBP2aA; PEBP2aA1; Polyomavirus enhancer binding protein 2 alpha A subunit; Polyomavirus enhancer-binding protein 2 alpha A subunit; Runt domain; Runt related transcription factor 2; Runt-related transcription factor 2; RUNX2; RUNX2_HUMAN; SL3 3 enhancer factor 1 alpha A subunit; SL3-3 enhancer factor 1 alpha A subunit; SL3/AKV core binding factor alpha A subunit; SL3/AKV core-binding factor alpha A subunit;

Immunogens

Immunogen:
Uniprot:

Q13950(RUNX2_HUMAN) >>Visit HPA database.

Gene(ID):
RUNX2(860)
Expression:
Q13950 RUNX2_HUMAN:

Specifically expressed in osteoblasts.

Sequence:
MASNSLFSTVTPCQQNFFWDPSTSRRFSPPSSSLQPGKMSDVSPVVAAQQQQQQQQQQQQQQQQQQQQQQQEAAAAAAAAAAAAAAAAAVPRLRPPHDNRTMVEIIADHPAELVRTDSPNFLCSVLPSHWRCNKTLPVAFKVVALGEVPDGTVVTVMAGNDENYSAELRNASAVMKNQVARFNDLRFVGRSGRGKSFTLTITVFTNPPQVATYHRAIKVTVDGPREPRRHRQKLDDSKPSLFSDRLSDLGRIPHPSMRVGVPPQNPRPSLNSAPSPFNPQGQSQITDPRQAQSSPPWSYDQSYPSYLSQMTSPSIHSTTPLSSTRGTGLPAITDVPRRISDDDTATSDFCLWPSTLSKKSQAGASELGPFSDPRQFPSISSLTESRFSNPRMHYPATFTYTPPVTSGMSLGMSATTHYHTYLPPPYPGSSQSQSGPFQTSSTPYLYYGTSSGSYQFPMVPGGDRSPSRMLPPCTTTSNGSTLLNPNLPNQNDGVDADGSHSSSPTVLNSSGRMDESVWRPY

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

PTMs - Q13950 As Substrate

Site PTM Type Enzyme
S24 Phosphorylation P27361 (MAPK3)
S28 Phosphorylation P31749 (AKT1) , P27361 (MAPK3)
S32 O-Glycosylation
S33 O-Glycosylation
S33 Phosphorylation
S43 Phosphorylation P27361 (MAPK3)
S118 Phosphorylation P45983 (MAPK8)
K176 Ubiquitination
S196 Phosphorylation P31749 (AKT1)
T198 Phosphorylation P31749 (AKT1)
T200 Phosphorylation P31749 (AKT1)
S237 Phosphorylation
K238 Acetylation
K238 Ubiquitination
S247 Phosphorylation
R258 Methylation
R267 Methylation
S275 Phosphorylation P27361 (MAPK3)
S294 Phosphorylation P27361 (MAPK3)
S312 Phosphorylation P27361 (MAPK3)
S314 Phosphorylation P27361 (MAPK3)
S340 Phosphorylation P27361 (MAPK3)
S347 Phosphorylation
S371 O-Glycosylation
R386 Methylation
S388 Phosphorylation
S451 Phosphorylation
S465 Phosphorylation P06493 (CDK1)
S480 Phosphorylation
S499 Phosphorylation
S503 Phosphorylation P27361 (MAPK3)
Y521 Phosphorylation

Research Backgrounds

Function:

Transcription factor involved in osteoblastic differentiation and skeletal morphogenesis. Essential for the maturation of osteoblasts and both intramembranous and endochondral ossification. CBF binds to the core site, 5'-PYGPYGGT-3', of a number of enhancers and promoters, including murine leukemia virus, polyomavirus enhancer, T-cell receptor enhancers, osteocalcin, osteopontin, bone sialoprotein, alpha 1(I) collagen, LCK, IL-3 and GM-CSF promoters. In osteoblasts, supports transcription activation: synergizes with SPEN/MINT to enhance FGFR2-mediated activation of the osteocalcin FGF-responsive element (OCFRE) (By similarity). Inhibits KAT6B-dependent transcriptional activation.

PTMs:

Phosphorylated; probably by MAP kinases (MAPK). Phosphorylation by HIPK3 is required for the SPEN/MINT and FGF2 transactivation during osteoblastic differentiation (By similarity). Phosphorylation at Ser-451 by CDK1 promotes endothelial cell proliferation required for tumor angiogenesis probably by facilitating cell cycle progression. Isoform 3 is phosphorylated on Ser-340.

Subcellular Location:

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:

Specifically expressed in osteoblasts.

Subunit Structure:

Heterodimer of an alpha and a beta subunit. The alpha subunit binds DNA as a monomer and through the Runt domain. DNA-binding is increased by heterodimerization. Interacts with XRCC6 (Ku70) and XRCC5 (Ku80). Interacts with HIVEP3. Interacts with IFI204. Interaction with SATB2; the interaction results in enhanced DNA binding and transactivation by these transcription factors. Binds to HIPK3. Interacts (isoform 3) with DDX5. Interacts with FOXO1 (via a C-terminal region); the interaction inhibits RUNX2 transcriptional activity towards BGLAP. This interaction is prevented on insulin or IGF1 stimulation as FOXO1 is exported from the nucleus (By similarity). Interacts with CCNB1, KAT6A and KAT6B. Interacts with FOXP3. Interacts with TMEM119 (By similarity). Interacts with OLFM2 (By similarity).

Family&Domains:

A proline/serine/threonine rich region at the C-terminus is necessary for transcriptional activation of target genes and contains the phosphorylation sites.

Research Fields

· Human Diseases > Cancers: Overview > Transcriptional misregulation in cancer.

References

1). LncRNA H19 Regulates BMP2-Induced Hypertrophic Differentiation of Mesenchymal Stem Cells by Promoting Runx2 Phosphorylation. Frontiers in Cell and Developmental Biology (PubMed: 32903671) [IF=5.5]

Application: IHC    Species: mice    Sample: flanks

Figure 5.. (D) The immunohistochemical staining was utilized to confirm the influence of silencing of H19 in BMP2-induced hypertrophic differentiation in vivo. The expression of Col10α1, MMP13, and Runx2 in the AdBMP2 group were less and weaken compared with the AdBMP2 + AdSimH19 group (a,b,c,d,e,f; a’,b’,c’,d’,e’,f’).
2). Polycystin‐1 modulates RUNX2 activation and osteocalcin gene expression via ERK signalling in a human craniosynostosis cell model. Journal of Cellular and Molecular Medicine (PubMed: 33656806) [IF=5.3]

Application: WB    Species: Human    Sample: trigonocephaly cranial suture cells

FIGURE 2 Effect of IgPKD1, MEK inhibitor and dual IgPKD1/MEK inhibitor treatment on phosphorylated RUNX2 (p‐RUNX2) in trigonocephaly cranial suture cells. A–C, Western blot analysis showing the effect of IgPKD1, MEK inhibitor and dual IgPKD1/MEK inhibitor treatment on the phosphorylation status of RUNX2 in trigonocephaly cells at 6, 12 and 24 h. D–F, Quantitative data showing the effect of IgPKD1, MEK inhibitor and dual IgPKD1/MEK inhibitor treatment on the phosphorylation status of RUNX2 in trigonocephaly cells at 6, 12 and 24 h. Bars represent means ± SD. ***P < .001
3). Enhanced Bone Regeneration through Regulation of Mechanoresponsive FAK-ERK1/2 Signaling by ZINC40099027 during Distraction Osteogenesis. International journal of medical sciences (PubMed: 38164350) [IF=3.6]

Application: WB    Species: Rat    Sample:

Figure 6 ZN27 promoted bone regeneration during distraction osteogenesis in rats. A Workflow of animal experiments. B, C Three-dimensional reconstructions (B) and longitudinal images (C) of micro-CT data of tibial distraction regenerates after consolidation for 2 and 4 weeks. For the three-dimensional reconstructions in each group, the left and right images were respectively the outside and inside views of the tibial distraction regenerates. D Quantitative analysis of BV/TV of the tibial distraction regenerates. n = 6 per group. E Representative images of H&E and Masson's trichrome staining in the central interzone of the tibial distraction regenerates. Red arrows, cartilaginous tissue. Dotted arrows, fibrous tissue. Black arrows, trabecular bone. Scale bar: 200 µm. F, G The fibrous tissue was stained with PSR (labeled with “F”), followed by quantitative analysis. Scale bar: 200 µm. n = 3 per group. H, I Immunohistochemical staining of chondrogenic marker COL2, followed by quantitative analysis. Scale bar: 200 µm. n = 3 per group. J, K Immunohistochemical staining of osteogenic marker OCN, followed by quantitative analysis. Scale bar: 200 µm. n = 3 per group. L, M Immunohistochemical staining of p-ERK and p-RUNX2 after consolidation for 2 weeks, followed by quantitative analysis. Scale bar: 200 µm. n = 3 per group. Abbreviations: ZN27, ZINC40099027; BV/TV, bone volume/tissue volume; H&E, hematoxylin-eosin; PSR, picrosirius red; COL2, collagen type II; OCN, osteocalcin; ERK1/2, extracellular signal-regulated kinase 1/2; RUNX2, runt-related transcription factor 2. Data are presented as the mean ± standard deviation. For D, G, I, and K, two-way ANOVA with Sidak's multiple comparisons test was used. For M, unpaired t test was employed.

Application: IF/ICC    Species: Rat    Sample:

Figure 6 ZN27 promoted bone regeneration during distraction osteogenesis in rats. A Workflow of animal experiments. B, C Three-dimensional reconstructions (B) and longitudinal images (C) of micro-CT data of tibial distraction regenerates after consolidation for 2 and 4 weeks. For the three-dimensional reconstructions in each group, the left and right images were respectively the outside and inside views of the tibial distraction regenerates. D Quantitative analysis of BV/TV of the tibial distraction regenerates. n = 6 per group. E Representative images of H&E and Masson's trichrome staining in the central interzone of the tibial distraction regenerates. Red arrows, cartilaginous tissue. Dotted arrows, fibrous tissue. Black arrows, trabecular bone. Scale bar: 200 µm. F, G The fibrous tissue was stained with PSR (labeled with “F”), followed by quantitative analysis. Scale bar: 200 µm. n = 3 per group. H, I Immunohistochemical staining of chondrogenic marker COL2, followed by quantitative analysis. Scale bar: 200 µm. n = 3 per group. J, K Immunohistochemical staining of osteogenic marker OCN, followed by quantitative analysis. Scale bar: 200 µm. n = 3 per group. L, M Immunohistochemical staining of p-ERK and p-RUNX2 after consolidation for 2 weeks, followed by quantitative analysis. Scale bar: 200 µm. n = 3 per group. Abbreviations: ZN27, ZINC40099027; BV/TV, bone volume/tissue volume; H&E, hematoxylin-eosin; PSR, picrosirius red; COL2, collagen type II; OCN, osteocalcin; ERK1/2, extracellular signal-regulated kinase 1/2; RUNX2, runt-related transcription factor 2. Data are presented as the mean ± standard deviation. For D, G, I, and K, two-way ANOVA with Sidak's multiple comparisons test was used. For M, unpaired t test was employed.

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