Product: Phospho-STAT3 (Ser727) Antibody
Catalog: AF3294
Description: Rabbit polyclonal antibody to Phospho-STAT3 (Ser727)
Application: WB IHC IF/ICC IP
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Chicken
Mol.Wt.: 86kDa; 88kD(Calculated).
Uniprot: P40763
RRID: AB_2834713

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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, IP 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(91%), Bovine(91%), Horse(91%), Sheep(82%), Rabbit(91%), Chicken(91%)
Clonality:
Polyclonal
Specificity:
Phospho-STAT3 (Ser727) Antibody detects endogenous levels of STAT3 only when phosphorylated at Serine 727.
RRID:
AB_2834713
Cite Format: Affinity Biosciences Cat# AF3294, RRID:AB_2834713.
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

1110034C02Rik; Acute Phase Response Factor; Acute-phase response factor; ADMIO; APRF; AW109958; DNA binding protein APRF; FLJ20882; HIES; MGC16063; Signal transducer and activator of transcription 3 (acute phase response factor); Signal transducer and activator of transcription 3; STAT 3; Stat3; STAT3_HUMAN;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P40763 STAT3_HUMAN:

Heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas.

Description:
The protein encoded by this gene is a member of the STAT protein family. In response to cytokines and growth factors, STAT family members are phosphorylated by the receptor associated kinases, and then form homo- or heterodimers that translocate to the cell nucleus where they act as transcription activators.
Sequence:
MAQWNQLQQLDTRYLEQLHQLYSDSFPMELRQFLAPWIESQDWAYAASKESHATLVFHNLLGEIDQQYSRFLQESNVLYQHNLRRIKQFLQSRYLEKPMEIARIVARCLWEESRLLQTAATAAQQGGQANHPTAAVVTEKQQMLEQHLQDVRKRVQDLEQKMKVVENLQDDFDFNYKTLKSQGDMQDLNGNNQSVTRQKMQQLEQMLTALDQMRRSIVSELAGLLSAMEYVQKTLTDEELADWKRRQQIACIGGPPNICLDRLENWITSLAESQLQTRQQIKKLEELQQKVSYKGDPIVQHRPMLEERIVELFRNLMKSAFVVERQPCMPMHPDRPLVIKTGVQFTTKVRLLVKFPELNYQLKIKVCIDKDSGDVAALRGSRKFNILGTNTKVMNMEESNNGSLSAEFKHLTLREQRCGNGGRANCDASLIVTEELHLITFETEVYHQGLKIDLETHSLPVVVISNICQMPNAWASILWYNMLTNNPKNVNFFTKPPIGTWDQVAEVLSWQFSSTTKRGLSIEQLTTLAEKLLGPGVNYSGCQITWAKFCKENMAGKGFSFWVWLDNIIDLVKKYILALWNEGYIMGFISKERERAILSTKPPGTFLLRFSESSKEGGVTFTWVEKDISGKTQIQSVEPYTKQQLNNMSFAEIIMGYKIMDATNILVSPLVYLYPDIPKEEAFGKYCRPESQEHPEADPGSAAPYLKTKFICVTPTTCSNTIDLPMSPRTLDSLMQFGNNGEGAEPSAGGQFESLTFDMELTSECATSPM

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
91
Horse
91
Bovine
91
Chicken
91
Rabbit
91
Sheep
82
Zebrafish
75
Dog
0
Xenopus
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P40763 As Substrate

Site PTM Type Enzyme
A2 Acetylation
S25 Phosphorylation
Y45 Phosphorylation
K49 Acetylation
K49 Methylation
Y68 Phosphorylation
Y79 Phosphorylation
K87 Acetylation
K87 Ubiquitination
K97 Ubiquitination
K140 Ubiquitination
K153 Ubiquitination
K161 Ubiquitination
K163 Ubiquitination
Y176 Phosphorylation
K177 Sumoylation
K177 Ubiquitination
K180 Ubiquitination
S181 Phosphorylation
S194 Phosphorylation
T196 Phosphorylation
K199 Ubiquitination
T236 Phosphorylation
K244 Ubiquitination
K283 Ubiquitination
K290 Ubiquitination
K294 Ubiquitination
K318 Ubiquitination
K348 Ubiquitination
K363 Ubiquitination
K365 Ubiquitination
K370 Acetylation
K370 Ubiquitination
K383 Ubiquitination
K409 Ubiquitination
S429 Phosphorylation
T440 Phosphorylation
Y446 Phosphorylation
K451 Sumoylation
K495 Sumoylation
Y539 Phosphorylation
S540 Phosphorylation
K551 Ubiquitination
K574 Ubiquitination
K601 Acetylation
K601 Ubiquitination
T605 Phosphorylation
K615 Acetylation
K615 Ubiquitination
K626 Ubiquitination
K631 Acetylation
K631 Ubiquitination
T632 Phosphorylation
S636 Phosphorylation
Y640 Phosphorylation
T641 Phosphorylation
K642 Ubiquitination
Y674 Phosphorylation
K679 Ubiquitination
K685 Acetylation
K685 Ubiquitination
Y686 Phosphorylation
S691 Phosphorylation
S701 Phosphorylation
Y705 Phosphorylation O60674 (JAK2) , Q15300 (RET/PTC2) , P23458 (JAK1) , P22455 (FGFR4) , P27361 (MAPK3) , Q13882 (PTK6) , P22607 (FGFR3) , Q9UM73 (ALK) , P14618 (PKM) , P16591 (FER) , P12931 (SRC)
K707 Acetylation
K707 Ubiquitination
T708 Phosphorylation
T714 Phosphorylation
T717 Phosphorylation
S719 Phosphorylation
T721 Phosphorylation
S727 Phosphorylation P51812 (RPS6KA3) , P45984-2 (MAPK9) , Q13233 (MAP3K1) , O75582 (RPS6KA5) , P28482 (MAPK1) , Q13555 (CAMK2G) , O43293 (DAPK3) , P42345 (MTOR) , P27361 (MAPK3) , Q02156 (PRKCE) , P45983 (MAPK8) , P51617 (IRAK1) , Q16539 (MAPK14) , Q05655 (PRKCD) , Q9UBE8 (NLK) , P06493 (CDK1) , Q00535 (CDK5) , Q9HC98 (NEK6)
S754 Phosphorylation

Research Backgrounds

Function:

Signal transducer and transcription activator that mediates cellular responses to interleukins, KITLG/SCF, LEP and other growth factors. Once activated, recruits coactivators, such as NCOA1 or MED1, to the promoter region of the target gene. May mediate cellular responses to activated FGFR1, FGFR2, FGFR3 and FGFR4. Binds to the interleukin-6 (IL-6)-responsive elements identified in the promoters of various acute-phase protein genes. Activated by IL31 through IL31RA. Acts as a regulator of inflammatory response by regulating differentiation of naive CD4(+) T-cells into T-helper Th17 or regulatory T-cells (Treg): deacetylation and oxidation of lysine residues by LOXL3, leads to disrupt STAT3 dimerization and inhibit its transcription activity. Involved in cell cycle regulation by inducing the expression of key genes for the progression from G1 to S phase, such as CCND1. Mediates the effects of LEP on melanocortin production, body energy homeostasis and lactation (By similarity). May play an apoptotic role by transctivating BIRC5 expression under LEP activation. Cytoplasmic STAT3 represses macroautophagy by inhibiting EIF2AK2/PKR activity. Plays a crucial role in basal beta cell functions, such as regulation of insulin secretion (By similarity).

PTMs:

Tyrosine phosphorylated upon stimulation with EGF. Tyrosine phosphorylated in response to constitutively activated FGFR1, FGFR2, FGFR3 and FGFR4 (By similarity). Activated through tyrosine phosphorylation by BMX. Tyrosine phosphorylated in response to IL6, IL11, LIF, CNTF, KITLG/SCF, CSF1, EGF, PDGF, IFN-alpha, LEP and OSM. Activated KIT promotes phosphorylation on tyrosine residues and subsequent translocation to the nucleus. Phosphorylated on serine upon DNA damage, probably by ATM or ATR. Serine phosphorylation is important for the formation of stable DNA-binding STAT3 homodimers and maximal transcriptional activity. ARL2BP may participate in keeping the phosphorylated state of STAT3 within the nucleus. Upon LPS challenge, phosphorylated within the nucleus by IRAK1. Upon erythropoietin treatment, phosphorylated on Ser-727 by RPS6KA5. Phosphorylation at Tyr-705 by PTK6 or FER leads to an increase of its transcriptional activity. Dephosphorylation on tyrosine residues by PTPN2 negatively regulates IL6/interleukin-6 signaling.

Acetylated on lysine residues by CREBBP. Deacetylation by LOXL3 leads to disrupt STAT3 dimerization and inhibit STAT3 transcription activity. Oxidation of lysine residues to allysine on STAT3 preferentially takes place on lysine residues that are acetylated.

Some lysine residues are oxidized to allysine by LOXL3, leading to disrupt STAT3 dimerization and inhibit STAT3 transcription activity. Oxidation of lysine residues to allysine on STAT3 preferentially takes place on lysine residues that are acetylated.

(Microbial infection) Phosphorylated on Tyr-705 in the presence of S.typhimurium SarA.

S-palmitoylated by ZDHHC19 in SH2 putative lipid-binding pockets, leading to homodimerization. Nuclear STAT3 is highly palmitoylated (about 75%) compared with cytoplasmic STAT3 (about 20%).

S-stearoylated, probably by ZDHHC19.

Subcellular Location:

Cytoplasm. Nucleus.
Note: Shuttles between the nucleus and the cytoplasm. Translocated into the nucleus upon tyrosine phosphorylation and dimerization, in response to signaling by activated FGFR1, FGFR2, FGFR3 or FGFR4. Constitutive nuclear presence is independent of tyrosine phosphorylation. Predominantly present in the cytoplasm without stimuli. Upon leukemia inhibitory factor (LIF) stimulation, accumulates in the nucleus. The complex composed of BART and ARL2 plays an important role in the nuclear translocation and retention of STAT3. Identified in a complex with LYN and PAG1.

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

Heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas.

Subunit Structure:

Forms a homodimer or a heterodimer with a related family member (at least STAT1). Interacts with IL31RA, NCOA1, PELP1, SIPAR, SOCS7, STATIP1 and TMF1 (By similarity). Interacts with IL23R in presence of IL23. Interacts (via SH2 domain) with NLK. Interacts with ARL2BP; the interaction is enhanced by LIF and JAK1 expression (By similarity). Interacts with KPNA4 and KPNA5; KPNA4 may be the primary mediator of nuclear import (By similarity). Interacts with CAV2; the interaction is increased on insulin-induced tyrosine phosphorylation of CAV2 and leads to STAT3 activation (By similarity). Interacts with ARL2BP; interaction is enhanced with ARL2. Interacts with NEK6 (By similarity). Binds to CDK9 when activated and nuclear. Interacts with BMX. Interacts with ZIPK/DAPK3. Interacts with PIAS3; the interaction occurs on stimulation by IL6, CNTF or OSM and inhibits the DNA binding activity of STAT3. In prostate cancer cells, interacts with STAT3 and promotes DNA binding activity of STAT3. Interacts with STMN3, antagonizing its microtubule-destabilizing activity. Interacts with the 'Lys-129' acetylated form of BIRC5/survivin. Interacts with FER. Interacts (via SH2 domain) with EIF2AK2/PKR (via the kinase catalytic domain). Interacts with STAT3; the interaction is independent of STAT3 Tyr-705 phosphorylation status. Interacts with FGFR4. Interacts with OCAD1 (By similarity). Interacts with ZDHHC19, leading to palmitoylation which promotes homodimerization and activation.

(Microbial infection) Interacts with HCV core protein.

(Microbial infection) Interacts with S.typhimurium SarA.

Family&Domains:

Belongs to the transcription factor STAT family.

Research Fields

· Cellular Processes > Cell growth and death > Necroptosis.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Signaling pathways regulating pluripotency of stem cells.   (View pathway)

· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > FoxO signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Jak-STAT signaling pathway.   (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > EGFR tyrosine kinase inhibitor resistance.

· Human Diseases > Endocrine and metabolic diseases > Insulin resistance.

· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.

· Human Diseases > Infectious diseases: Viral > Hepatitis C.

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Infectious diseases: Viral > Measles.

· Human Diseases > Infectious diseases: Viral > Epstein-Barr virus infection.

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Viral carcinogenesis.

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Cancers: Overview > MicroRNAs in cancer.

· Human Diseases > Cancers: Specific types > Pancreatic cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Acute myeloid leukemia.   (View pathway)

· Human Diseases > Cancers: Specific types > Non-small cell lung cancer.   (View pathway)

· Human Diseases > Immune diseases > Inflammatory bowel disease (IBD).

· Organismal Systems > Immune system > Chemokine signaling pathway.   (View pathway)

· Organismal Systems > Immune system > Th17 cell differentiation.   (View pathway)

· Organismal Systems > Endocrine system > Prolactin signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Adipocytokine signaling pathway.

References

1). Mesenchymal Stem Cells Overexpressing ACE2 Favorably Ameliorate LPS-Induced Inflammatory Injury in Mammary Epithelial Cells. Frontiers in Immunology (PubMed: 35095873) [IF=7.3]

Application: WB    Species: Mice    Sample: EpH4-Ev cells

Figure 7 MSC-ACE2 upregulates IL-10/STAT3/SOCS3 signaling pathway expression to suppress LPS-induced inflammation in EpH4-Ev cells. (A) Detection of relative protein expression levels of IL-10, phosphorylation levels of STAT3, STAT3 and SOCS3 by Western blot; (B–D), Statistics of the IL-10, phosphorylation levels of STAT3, STAT3 and SOCS3 Western blot results. Experiments were repeated three times and data were presented as the mean ± SEM (n = 4). * P < 0.05 vs. EpH4-Ev; # P < 0.05 vs. LPS; $ P < 0.05 vs. MSC; & P < 0.05 vs. MSC-GFP.

2). Cyclin G2 inhibits oral squamous cell carcinoma growth and metastasis by binding to IGFBP3 and regulating the FAK-SRC-STAT signaling pathway. Frontiers in Oncology (PubMed: 33240810) [IF=4.7]

Application: WB    Species: Human    Sample: SCC-9 cells

Figure 5 Cyclin G2 inhibits the FAK-SRC-STAT pathway in vitro and in vivo. (A) The protein expressions along with quantification of FAK, P-FAK, SRC, P-SRC, STAT3, P-STAT3, Bcl-2, c-Myc, and MMP9 in the FAK-SRC-STAT pathway. (B–D) The relative mRNA expressions of Bcl-2, c-Myc and MMP9. (E) Analysis of MMP9 secretion and its quantification in SCC-9 cells. (F, G) Immunohistochemical staining and analysis of the expression of cyclin G2, p-FAK, p-SRC and p-STAT3. Scale bar = 100 µm. *p < 0.05, **p < 0.01, ***p < 0.001 vs. vector.

3). Qing-Luo-Yin Alleviated Experimental Arthritis in Rats by Disrupting Immune Feedback Between Inflammatory T Cells and Monocytes: Key Evidences from Its Effects on Immune Cell Phenotypes. Journal of Inflammation Research (PubMed: 35002280) [IF=4.5]

4). Umbilical cord‑derived mesenchymal stem cells can inhibit the biological functions of melanoma A375 cells. ONCOLOGY REPORTS (PubMed: 29767256) [IF=4.2]

5). LYPD8 regulates the proliferation and migration of colorectal cancer cells through inhibiting the secretion of IL‑6 and TNF‑α. Oncology Reports (PubMed: 30816524) [IF=4.2]

6). STAT3/miR-15a-5p/CX3CL1 Loop Regulates Proliferation and Migration of Vascular Endothelial Cells in Atherosclerosis. International Journal of Medical Sciences (PubMed: 33456354) [IF=3.6]

Application: WB    Species: Human    Sample: HUVECs

Figure 2 CX3CL1 expression activated in HUVECs by STAT3 signaling pathway. A-D: The effect of treated with IL-6 on CX3CL1 expression in HUVECs was detected by western blot analysis and qRT-PCR. Cells were stimulated by IL-6 at various concentrations (0, 10, 20, 40 60 and 80, 100 ng/mL) for 24 hours or at different times (0, 6, 12, 24, and 48h) at a concentration of 80ng/mL. All experiments were repeated three times in duplicate, and the semi-quantitative analysis was calculated by SPSS statistical soft. (**P< 0.01). E and F: The effect of treated with overexpression STAT3 or knocked down STAT3 on the expression of CX3CL1 was examined by western blot analysis and qRT-PCR in HUVECs. All experiments were repeated three times in duplicate, and the semi-quantitative analysis was calculated by SPSS statistical soft. (**P< 0.01)

7). miR-142-5p Inhibits Cell Invasion and Migration by Targeting DNMT1 in Breast Cancer. Oncology Research (PubMed: 34321149) [IF=3.1]

8). Biological functions of lung cancer cells are suppressed in co-culture with mesenchymal stem cells isolated from umbilical cord. Experimental and Therapeutic Medicine (PubMed: 29399109) [IF=2.7]

9). Cyclovirobuxine D pretreatment ameliorates septic heart injury through mitigation of ferroptosis. Experimental and Therapeutic Medicine (PubMed: 37522059) [IF=2.7]

Application: WB    Species: Rat    Sample: H9C2 cells

Figure 2 CVB-D inhibits cellular iron-overload induced by LPS. (A) Calcein-AM imaging of intracellular iron levels in H9C2 cells. (B) Western blot images and protein expression levels of DMT1, TfR1, FPN1 and FtH in LPS- and CVB-D-treated H9C2 cells. (C) hamp mRNA expression levels in LPS- and CVB-D-treated H9C2 cells. (D) Protein expression levels of pro-IL-1β, TNF-α, IL-6, p-STAT3 and STAT3 in LPS- and CVB-D-treated H9C2 cells. Data are presented as the mean ± SD (n=6). **P

10). Huang Gan Formula Eliminates the Oxidative Stress Effects of Advanced Oxidation Protein Products on the Divergent Regulation of the Expression of AGEs Receptors via the JAK2/STAT3 Pathway. Evidence-based Complementary and Alternative Medicine (PubMed: 28465704)

Application: WB    Species: human    Sample:

Figure 6: Effects of AOPPs on JAK2 and STAT3 phosphorylation. Serum-starved HMCs were incubated with 200

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