Product: Phospho-STAT1 (Tyr701) Antibody
Catalog: AF3300
Source: Rabbit
Application: WB, IHC, IF/ICC, IP, ELISA(peptide)
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.: 84kD; 87kD(Calculated).
Uniprot: P42224
RRID: AB_2834719

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IP, IF/ICC 1:100-1:500, ELISA(peptide) 1:20000-1:40000
*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%), Rabbit(100%), Dog(100%), Chicken(80%), Xenopus(100%)
Clonality:
Polyclonal
Specificity:
Phospho-STAT1 (Tyr701) Antibody detects endogenous levels of STAT1 only when phosphorylated at Tyrosine 701.
RRID:
AB_2834719
Cite Format: Affinity Biosciences Cat# AF3300, RRID:AB_2834719.
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

CANDF7; DKFZp686B04100; ISGF 3; ISGF3; OTTHUMP00000163552; OTTHUMP00000165046; OTTHUMP00000165047; OTTHUMP00000205845; Signal transducer and activator of transcription 1; Signal transducer and activator of transcription 1, 91kDa; Signal transducer and activator of transcription 1-alpha/beta; Stat1; STAT1_HUMAN; STAT91; Transcription factor ISGF-3 components p91/p84;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
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:
MSQWYELQQLDSKFLEQVHQLYDDSFPMEIRQYLAQWLEKQDWEHAANDVSFATIRFHDLLSQLDDQYSRFSLENNFLLQHNIRKSKRNLQDNFQEDPIQMSMIIYSCLKEERKILENAQRFNQAQSGNIQSTVMLDKQKELDSKVRNVKDKVMCIEHEIKSLEDLQDEYDFKCKTLQNREHETNGVAKSDQKQEQLLLKKMYLMLDNKRKEVVHKIIELLNVTELTQNALINDELVEWKRRQQSACIGGPPNACLDQLQNWFTIVAESLQQVRQQLKKLEELEQKYTYEHDPITKNKQVLWDRTFSLFQQLIQSSFVVERQPCMPTHPQRPLVLKTGVQFTVKLRLLVKLQELNYNLKVKVLFDKDVNERNTVKGFRKFNILGTHTKVMNMEESTNGSLAAEFRHLQLKEQKNAGTRTNEGPLIVTEELHSLSFETQLCQPGLVIDLETTSLPVVVISNVSQLPSGWASILWYNMLVAEPRNLSFFLTPPCARWAQLSEVLSWQFSSVTKRGLNVDQLNMLGEKLLGPNASPDGLIPWTRFCKENINDKNFPFWLWIESILELIKKHLLPLWNDGCIMGFISKERERALLKDQQPGTFLLRFSESSREGAITFTWVERSQNGGEPDFHAVEPYTKKELSAVTFPDIIRNYKVMAAENIPENPLKYLYPNIDKDHAFGKYYSRPKEAPEPMELDGPKGTGYIKTELISVSEVHPSRLQTTDNLLPMSPEEFDEVSRIVGSVEFDSMMNTV

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

PTMs - P42224 As Substrate

Site PTM Type Enzyme
S2 Acetylation
S2 Phosphorylation
R31 Methylation
K40 Ubiquitination
Y68 Phosphorylation
Y106 Phosphorylation
K114 Methylation
K114 Sumoylation
K114 Ubiquitination
K138 Ubiquitination
K140 Ubiquitination
K145 Ubiquitination
K152 Ubiquitination
K161 Ubiquitination
S162 Phosphorylation
Y170 Phosphorylation
K173 Acetylation
K173 Ubiquitination
K175 Methylation
K175 Ubiquitination
T184 Phosphorylation
K193 Sumoylation
K193 Ubiquitination
K200 Ubiquitination
K201 Acetylation
K201 Ubiquitination
K209 Ubiquitination
K240 Ubiquitination
K286 Ubiquitination
K296 Methylation
K296 Ubiquitination
K298 Ubiquitination
K350 Ubiquitination
K359 Ubiquitination
K361 Ubiquitination
K366 Methylation
K366 Ubiquitination
T373 Phosphorylation
K375 Ubiquitination
K379 Ubiquitination
K388 Ubiquitination
K410 Acetylation
K410 Ubiquitination
K413 Acetylation
T417 Phosphorylation
K511 Ubiquitination
K525 Methylation
K525 Ubiquitination
S532 Phosphorylation
K544 Ubiquitination
S583 Phosphorylation
K592 Ubiquitination
K636 Ubiquitination
K637 Methylation
K637 Ubiquitination
S640 Phosphorylation
K652 Sumoylation
K652 Ubiquitination
K665 Methylation
K665 Ubiquitination
Y668 Phosphorylation
K673 Ubiquitination
K679 Sumoylation
K679 Ubiquitination
K685 Ubiquitination
K697 Ubiquitination
T699 Phosphorylation
Y701 Phosphorylation P52333 (JAK3) , P22455 (FGFR4) , P23458 (JAK1) , P22607 (FGFR3) , P00533 (EGFR) , O60674 (JAK2) , P12931 (SRC)
K703 Sumoylation
K703 Ubiquitination
T704 Phosphorylation
S708 Phosphorylation
S710 Phosphorylation
S715 Phosphorylation
S727 Phosphorylation Q13554 (CAMK2B) , P49336 (CDK8) , Q05655 (PRKCD) , P68400 (CSNK2A1) , Q14164 (IKBKE) , Q15759 (MAPK11) , P19525 (EIF2AK2) , P27361 (MAPK3) , P28482 (MAPK1) , P45983 (MAPK8) , P45984 (MAPK9) , O75582 (RPS6KA5) , Q16539 (MAPK14)
S740 Phosphorylation
S745 Phosphorylation
T749 Phosphorylation

Research Backgrounds

Function:

Signal transducer and transcription activator that mediates cellular responses to interferons (IFNs), cytokine KITLG/SCF and other cytokines and other growth factors. Following type I IFN (IFN-alpha and IFN-beta) binding to cell surface receptors, signaling via protein kinases leads to activation of Jak kinases (TYK2 and JAK1) and to tyrosine phosphorylation of STAT1 and STAT2. The phosphorylated STATs dimerize and associate with ISGF3G/IRF-9 to form a complex termed ISGF3 transcription factor, that enters the nucleus. ISGF3 binds to the IFN stimulated response element (ISRE) to activate the transcription of IFN-stimulated genes (ISG), which drive the cell in an antiviral state. In response to type II IFN (IFN-gamma), STAT1 is tyrosine- and serine-phosphorylated. It then forms a homodimer termed IFN-gamma-activated factor (GAF), migrates into the nucleus and binds to the IFN gamma activated sequence (GAS) to drive the expression of the target genes, inducing a cellular antiviral state. Becomes activated in response to KITLG/SCF and KIT signaling. May mediate cellular responses to activated FGFR1, FGFR2, FGFR3 and FGFR4.

PTMs:

Phosphorylated on tyrosine and serine residues in response to a variety of cytokines/growth hormones including IFN-alpha, IFN-gamma, PDGF and EGF. Activated KIT promotes phosphorylation on tyrosine residues and subsequent translocation to the nucleus. Upon EGF stimulation, phosphorylation on Tyr-701 (lacking in beta form) by JAK1, JAK2 or TYK2 promotes dimerization and subsequent translocation to the nucleus. Growth hormone (GH) activates STAT1 signaling only via JAK2. Tyrosine phosphorylated in response to constitutively activated FGFR1, FGFR2, FGFR3 and FGFR4. Phosphorylation on Ser-727 by several kinases including MAPK14, ERK1/2 and CAMKII on IFN-gamma stimulation, regulates STAT1 transcriptional activity. Phosphorylation on Ser-727 promotes sumoylation though increasing interaction with PIAS. Phosphorylation on Ser-727 by PRKCD induces apoptosis in response to DNA-damaging agents. Phosphorylated on tyrosine residues when PTK2/FAK1 is activated; most likely this is catalyzed by a SRC family kinase. Dephosphorylation on tyrosine residues by PTPN2 negatively regulates interferon-mediated signaling. Upon viral infection or IFN induction, phosphorylation on Ser-708 occurs much later than phosphorylation on Tyr-701 and is required for the binding of ISGF3 on the ISREs of a subset of IFN-stimulated genes IKBKE-dependent. Phosphorylation at Tyr-701 and Ser-708 are mutually exclusive, phosphorylation at Ser-708 requires previous dephosphorylation of Tyr-701.

Sumoylated with SUMO1, SUMO2 and SUMO3. Sumoylation is enhanced by IFN-gamma-induced phosphorylation on Ser-727, and by interaction with PIAS proteins. Enhances the transactivation activity.

ISGylated.

Mono-ADP-ribosylated at Glu-657 and Glu-705 by PARP14; ADP-ribosylation prevents phosphorylation at Tyr-701. However, the role of ADP-ribosylation in the prevention of phosphorylation has been called into question and the lack of phosphorylation may be due to sumoylation of Lys-703.

Monomethylated at Lys-525 by SETD2; monomethylation is necessary for phosphorylation at Tyr-701, translocation into the nucleus and activation of the antiviral defense.

Subcellular Location:

Cytoplasm. Nucleus.
Note: Translocated into the nucleus upon tyrosine phosphorylation and dimerization, in response to IFN-gamma and signaling by activated FGFR1, FGFR2, FGFR3 or FGFR4 (PubMed:15322115). Monomethylation at Lys-525 is required for phosphorylation at Tyr-701 and translocation into the nucleus (PubMed:28753426). Translocates into the nucleus in response to interferon-beta stimulation (PubMed:26479788).

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

Isoform alpha homodimerizes upon IFN-gamma induced phosphorylation. Heterodimer with STAT2 upon IFN-alpha/beta induced phosphorylation. The heterodimer STAT1:STAT2 forms the interferon-stimulated gene factor 3 complex (ISGF3) with IRF9 (By similarity). Interacts (phosphorylated at Ser-727) with PIAS1; the interaction results in release of STAT1 from its target gene. Interacts with IFNAR1; the interaction requires the phosphorylation of IFNAR1 at 'Tyr-466'. Interacts with IFNAR2. Found in a complex with NMI and CREBBP/CBP. Interacts with NMI which is required for CREBBP/CBP recruitment to the complex. Interacts with PTK2/FAK1. Interacts with SRC (By similarity). Interacts with ERBB4 (phosphorylated). Interacts with PARP9 and DTX3L independently of IFN-beta or IFN-gamma-mediated STAT1 'Tyr-701' phosphorylation. Interacts with histone acetyltransferase EP300/p300 in response to INF-gamma stimulation. Interacts with OTOP1 (By similarity).

(Microbial infection) Interacts with Sendai virus C', C, Y1 and Y2 proteins, preventing activation of ISRE and GAS promoter.

(Microbial infection) Interacts with Nipah virus P, V and W proteins preventing activation of ISRE and GAS promoter.

(Microbial infection) Interacts with Rabies virus phosphoprotein preventing activation of ISRE and GAS promoter.

(Microbial infection) Interacts with HCV core protein; the interaction results in STAT1 degradation.

(Microbial infection) Interacts with ebolavirus protein VP24.

Family&Domains:

Belongs to the transcription factor STAT family.

Research Fields

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

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

· Human Diseases > Infectious diseases: Parasitic > Leishmaniasis.

· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.

· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.

· 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 > Influenza A.

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Infectious diseases: Viral > Herpes simplex infection.

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

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

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

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

· Organismal Systems > Development > Osteoclast differentiation.   (View pathway)

· Organismal Systems > Immune system > Toll-like receptor signaling pathway.   (View pathway)

· Organismal Systems > Immune system > NOD-like receptor signaling pathway.   (View pathway)

· Organismal Systems > Immune system > Th1 and Th2 cell differentiation.   (View pathway)

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

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

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

References

1). Sun Y et al. Chemical structure and anti-inflammatory activity of a branched polysaccharide isolated from Phellinus baumii. Carbohydr Polym 2021 Sep 15;268:118214. (PubMed: 34127216) [IF=10.723]

Application: WB    Species: Mouse    Sample: RAW 264.7 cells

Fig. 5. |Effects of SHPS-1 on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. The internal panels represent the effect of SHPS-1 and LPS on (A) cell proliferation, (B) nitric oxide concentration in cell culture medium, (C) gene expression, (D) proteins levels analyzed by western blot.

Application: IF/ICC    Species: Mouse    Sample: RAW 264.7 cells

Fig. 5. |Effects of SHPS-1 on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. The internal panels represent the effect of SHPS-1 and LPS on (A) cell proliferation, (B) nitric oxide concentration in cell culture medium, (C) gene expression, (D) proteins levels analyzed by western blot.(E) NF-KB p65 nucleus translocation analyzed by immunofluorescence (IF), (F) phosphorylation level of STAT1 analyzed by IF

2). Liu C et al. STAT1-mediated inhibition of FOXM1 enhances gemcitabine sensitivity in pancreatic cancer. Clin Sci (Lond) 2019 Feb 19 (PubMed: 30782607) [IF=6.876]

Application: WB    Species: human    Sample: SW1990 cells

Figure 6 |STAT1 directly suppresses FOXM1 expression in pancreatic cancer cells. (C) SW1990 cells were treated with IFNγ (100 ng/mL) and/or Fludarabine (10 μM) for 24h and then whole cell lysates were extracted. FOXM1, STAT1, and p STAT1 levels were analyzed using western blotting. The protein levels in each well were determined quantitatively by densitometry analysis (right panels). The experiments were performed three times.

Application: IHC    Species: human    Sample: pancreatic xenograft tumors

Figure 8 |IFNγ inhibits FOXM1 sensitization to gemcitabine in pancreatic xenograft tumors.(C) The representative tumor tissue sections from xenografts in different treatment groups, analyzed by immunohistochemistry for the expression of FOXM1, pSTAT1, the proliferation markers PCNA and apoptotic marker Cleaved Caspase3. Scale bar, 20 μm.

3). Liu Z et al. Guizhi Fuling pill attenuates liver fibrosis in vitro and in vivo via inhibiting TGF-β1/Smad2/3 and activating IFN-γ/Smad7 signaling pathways. Bioengineered 2022 Apr;13(4):9357-9368. (PubMed: 35387552) [IF=6.832]

Application: WB    Species: human    Sample: LX-2 cells

Figure 2.| GZFL inhibits acetaldehyde-induced LX-2 cells activation through suppressing TGF-β1/Smad2/3 signaling and activating IFN-γ/STAT1/Smad7 signaling. LX-2 cells were exposed to acetaldehyde (AA; 400 μM) for 24 h, followed by exposure to colchicine (4 μg/ml) or GZFL (8 or 10 mg/ml) for another 24 h. (a, b) TGF-β1, TGF-βR2, CUGBP1, p-STAT1, p-Smad2, p-Smad3, Smad7,α-SMA and Collagen I expressions in LX-2 cells were detected by western blot assay.

4). Wang J et al. Carotid baroreceptor stimulation improves cardiac performance and reverses ventricular remodelling in canines with pacing-induced heart failure. Life Sci 2019 Feb 24 (PubMed: 30811965) [IF=6.780]

5). Luo N et al. Diosmetin Ameliorates Nonalcoholic Steatohepatitis through Modulating Lipogenesis and Inflammatory Response in a STAT1/CXCL10-Dependent Manner. J Agric Food Chem 2021 Jan 20;69(2):655-667. (PubMed: 33404223) [IF=5.895]

Application: WB    Species: Mice    Sample: liver tissue

Figure 3. Effect of Dios on molecular phenotype of STAT1 and CXCL10 in mice liver by using RNA-Seq. (A−B) Identification of DEGs in different groups. (A) HFD vs SD group. (B) HFD + Dios vs HFD group. The red points represent upregulated genes. The green points represent downregulated genes. The blue points represent genes with no significant difference. DEGs were screened on P < 0.05 and | log2 (Fold change) | > 1. (C) Heat map analysis was employed to the discrimination of expression profile of DEGs across the samples. Red and blue areas separately represent highly and lowly expressed genes in mice livers among SD, HFD, and the HFD + Dios group. (D) Venn diagram of DEGs among different groups. 456 shared DEGs were obtained from this diagram. (E) The PPI network of 456 shared DEGs was analyzed through the STING database. There were 408 nodes and 3294 edged in the PPI network. (F) The ten hub genes including CXCL10 and STAT1 were confirmed by Cytohubba. (G) The expression of DEGs mRNA (FPR2, PSMB8, PTAFR, CCR7, CXCR4, and LCK) was measured under the method of quantitative RT-PCR. (H) The mRNA expression of STAT1 and CXCL10 in mice livers by qRT-PCR assay. (I) The expression of CXCL10, total STAT1, p-STAT1Y701, and p-STAT1S727 protein was examined through Western blot assay. β-Actin was used as an internal control. Data are expressed as mean ± SD (n = 3). *P < 0.05, vs the SD group; # P < 0.05, vs the HFD group.

6). Liu X et al. IRG1 increases MHC class I level in macrophages through STAT-TAP1 axis depending on NADPH oxidase mediated reactive oxygen species. Int Immunopharmacol 2017 Jul;48:76-83 (PubMed: 28477473) [IF=5.714]

Application: WB    Species: human    Sample:

Fig. 5. Activated STAT1/3 participates in ROS-mediated expression of TAP1 and PSMB9 in IRG1-overexpressed cells. (A) Luciferase reporter assay to analyze the relationship between ISRE and IRG1; (B) IRG1 activates the phosphorylation of STAT1 and STAT3 in different time; (C) Western blot analysis of TAP1 level after pretreating STAT1 or STAT3 inhibitors in IRG1-overexpressed macrophages; (D) Western blot analysis of TAP1 level after STAT1 or STAT3 knockdown in IRG1-overexpressed macrophages; (E) Flow cytometry analysis of MHC I level after STAT1 or STAT3 knockdown in IRG1-overexpressed macrophages (#: not significant vs. shNC group). (F) Western blot analysis of phosphorylation of STAT1 and STAT3 level after pretreating different antioxidant in IRG1-overexpressed macrophages. (*p < 0.05; **p < 0.01; ns: not significant, n = 3.)

7). Que ZJ et al. Proteomics analysis of tumor exosomes reveals vital pathways of Jinfukang inhibiting circulating tumor cells metastasis in lung cancer. J Ethnopharmacol 2020 Jun 28;256:112802 (PubMed: 32240782) [IF=5.195]

Application: WB    Species: human    Sample: CTC-TJH-01 cells

Fig. 6. |Jinfukang inhibits the migration of CTC cells through EGF signaling pathway. (A) CTC-TJH-01 cells were treated with Jinfukang (700 μg/mL) for 24 h. Protein level of EGF, p-mTOR, p-STAT1, STAT3, p-STAT3, PRKCA, p-PRKCA, Akt1/2/3 and p-Akt were analyzed by Western blot analysis.

8). Gao B et al. Knockdown of ISOC1 inhibits the proliferation and migration and induces the apoptosis of colon cancer cells through the AKT/GSK-3β pathway. Carcinogenesis 2019 Nov 19 (PubMed: 31740942) [IF=4.741]

9). Zhang Y et al. Dihydrotanshinone I Alleviates Crystalline Silica-Induced Pulmonary Inflammation by Regulation of the Th Immune Response and Inhibition of STAT1/STAT3. Mediators Inflamm 2019 Jul 9;2019:3427053 (PubMed: 31379467) [IF=4.529]

10). Min Zhao et al. HuoXueTongFu Formula Alleviates Intraperitoneal Adhesion by Regulating Macrophage Polarization and the SOCS/JAK2/STAT/PPAR-γ Signalling Pathway. MEDIAT INFLAMM 2019, Article ID 1769374, 17 pages [IF=4.529]

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