Product nameAnti-STAT1 (phospho Y701) antibody
See all STAT1 primary antibodies
DescriptionRabbit polyclonal to STAT1 (phospho Y701)
Specificityab30645 detects endogenous levels of STAT1 only when phosphorylated at Tyrosine 701.
Tested applicationsSuitable for: ICC, ELISA, IP, IHC-P, WB, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Human
Synthetic phosphopeptide derived from human STAT1 around the phosphorylation site of Tyrosine 701.
- IHC: Breast carcinoma tissue. WB: MC7 cells treated with EGF.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 50% Glycerol, 0.87% Sodium chloride
Without Mg2+ and Ca2+
Concentration information loading...
PurityImmunogen affinity purified
Purification notesThe antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific phosphopeptide. The antibody against non-phosphopeptide was removed by chromatography using non-phosphopeptide corresponding to the phosphorylation site.
Corresponding non-phospho antibody
Our Abpromise guarantee covers the use of ab30645 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration.|
|WB||1/500 - 1/1000. Detects a band of approximately 87 kDa (predicted molecular weight: 87 kDa).|
|ICC/IF||Use a concentration of 1 - 5 µg/ml.|
FunctionSignal transducer and activator of transcription that mediates signaling by interferons (IFNs). Following type I IFN (IFN-alpha and IFN-beta) binding to cell surface receptors, Jak kinases (TYK2 and JAK1) are activated, leading to tyrosine phosphorylation of STAT1 and STAT2. The phosphorylated STATs dimerize, 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 interferon stimulated genes, 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.
Involvement in diseaseNote=STAT1 deficiency results in impaired immune response leading to severe mycobacterial and viral diseases. In the case of complete deficiency, patients can die of viral disease.
Defects in STAT1 are a cause of mendelian susceptibility to mycobacterial disease (MSMD) [MIM:209950]; also known as familial disseminated atypical mycobacterial infection. This rare condition confers predisposition to illness caused by moderately virulent mycobacterial species, such as Bacillus Calmette-Guerin (BCG) vaccine and environmental non-tuberculous mycobacteria, and by the more virulent Mycobacterium tuberculosis. Other microorganisms rarely cause severe clinical disease in individuals with susceptibility to mycobacterial infections, with the exception of Salmonella which infects less than 50% of these individuals. The pathogenic mechanism underlying MSMD is the impairment of interferon-gamma mediated immunity whose severity determines the clinical outcome. Some patients die of overwhelming mycobacterial disease with lepromatous-like lesions in early childhood, whereas others develop, later in life, disseminated but curable infections with tuberculoid granulomas. MSMD is a genetically heterogeneous disease with autosomal recessive, autosomal dominant or X-linked inheritance.
Sequence similaritiesBelongs to the transcription factor STAT family.
Contains 1 SH2 domain.
modificationsPhosphorylated on tyrosine and serine residues in response to IFN-alpha, IFN-gamma, PDGF and EGF. Phosphorylation on Tyr-701 (lacking in beta form) by JAK promotes dimerization and subsequent translocation to the nucleus. 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 PKCdelta induces apoptosis in response to DNA-damaging agents.
Sumoylated by 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.
Cellular localizationCytoplasm. Nucleus. Translocated into the nucleus in response to IFN-gamma-induced tyrosine phosphorylation and dimerization.
- Information by UniProt
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All lanes : Anti-STAT1 (phospho Y701) antibody (ab30645)
Lane 1 : Untreated MCF7 lysate (5-30ug).
Lane 2 : EGF treated MCF7 lysate (5-30ug).
Predicted band size: 87 kDa
Observed band size: 87 kDa
Additional bands at: 47 kDa. We are unsure as to the identity of these extra bands.
Suggested dilution: 1:500 - 1:1000
ICC/IF image of ab30645 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab30645, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Paraffin-embedded human breast carcinoma tissue stained for STAT1 (phospho Y701) using ab30645 at 1/590 dilution in immunohistochemical analysis. Tissue was incubated in the absence (left) or presence (right) of immunizing phospho-peptide.
ab30645 staining STAT1 (phospho Y701) in murine intestine tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at 23°C; antigen retrieval was by heat mediation with a citrate buffer. Samples were incubated with primary antibody (1/400 in Tris Buffer Saline) for 16 hours at 4°C. An AlexaFluor®488-conjugated goat anti-rabbit polyclonal IgG (1/200) was used as the secondary antibody.
This product has been referenced in:
- Jiang L et al. The Pros1/Tyro3 axis protects against periodontitis by modulating STAT/SOCS signalling. J Cell Mol Med 23:2769-2781 (2019). Read more (PubMed: 30729671) »
- An JC et al. miR-944 inhibits lung adenocarcinoma tumorigenesis by targeting STAT1 interaction. Oncol Lett 17:3790-3798 (2019). Read more (PubMed: 30881499) »