Recombinant
RabMAb

Recombinant Anti-STAT1 antibody [EPR23049-111] - ChIP Grade (ab239360)

Overview

  • Product name

    Anti-STAT1 antibody [EPR23049-111] - ChIP Grade
    See all STAT1 primary antibodies
  • Description

    Rabbit monoclonal [EPR23049-111] to STAT1 - ChIP Grade
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt, ICC/IF, WB, IPmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fragment within Mouse STAT1 aa 550 to the C-terminus. The exact sequence is proprietary.
    Database link: P42225

  • Positive control

    • WB: Human lymph node, RAW264.7, HeLa, MCF7, Mouse lung, Mouse breast and NIH/3T3 lysates. ICC/IF: HeLa and RAW 264.7 cells. FC: RAW264.7 cells. IP: RAW264.7 cells.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab239360 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt 1/600.

 

 

ICC/IF 1/50.

 

 

WB 1/1000. Predicted molecular weight: 87 kDa.

 

 

IP 1/30.

 

 

  • Application notes
    Is unsuitable for IHC-P.
  • Target

    • Function

      Signal 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 disease

      Note=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 similarities

      Belongs to the transcription factor STAT family.
      Contains 1 SH2 domain.
    • Post-translational
      modifications

      Phosphorylated 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.
      ISGylated.
    • Cellular localization

      Cytoplasm. Nucleus. Translocated into the nucleus in response to IFN-gamma-induced tyrosine phosphorylation and dimerization.
    • Information by UniProt
    • Database links

    • Alternative names

      • Signal transducer and activator of transcription 1 91kD antibody
      • CANDF7 antibody
      • DKFZp686B04100 antibody
      • IMD31A antibody
      • IMD31B antibody
      • IMD31C antibody
      • ISGF 3 antibody
      • ISGF-3 antibody
      • OTTHUMP00000163552 antibody
      • OTTHUMP00000165046 antibody
      • OTTHUMP00000165047 antibody
      • OTTHUMP00000205845 antibody
      • Signal transducer and activator of transcription 1 91kDa antibody
      • Signal transducer and activator of transcription 1 antibody
      • Signal transducer and activator of transcription 1, 91kD antibody
      • Signal transducer and activator of transcription 1-alpha/beta antibody
      • STAT 1 antibody
      • Stat1 antibody
      • STAT1_HUMAN antibody
      • STAT91 antibody
      • Transcription factor ISGF 3 components p91 p84 antibody
      • Transcription factor ISGF-3 components p91/p84 antibody
      • Transcription factor ISGF3 components p91/p84 antibody
      • XStat1 antibody
      see all

    Images

    • Chromatin was prepared from HT-1080 cells treated with IFN-γ (50ng/ml 30min) according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.

      The  ChIP was performed with 25 µg of chromatin, 5 µg of  ab239360 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 20 µl of Protein A/G sepharose beads.  The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).

      Primers and probes are commercial primers from Cell Signaling Technology (Cat. No.: #5148)

      *http://www.abcam.com/resources?keywords=X%20ChIP%20protocol

    • All lanes : Anti-STAT1 antibody [EPR23049-111] - ChIP Grade (ab239360) at 1/1000 dilution

      Lane 1 : Human lymph node tissue lysate
      Lane 2 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate
      Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
      Lane 4 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate
      Lane 5 : Mouse lung tissue lysate
      Lane 6 : Mouse breast tissue lysate
      Lane 7 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 87 kDa
      Observed band size: 88 kDa
      why is the actual band size different from the predicted?



      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

      Lanes 5-7 were developed using a higher sensitivity ECL substrate. 

      Exposure time: Lanes 1-4: 3 minutes; Lane 5-7: 48 seconds.

    • Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling STAT1 with ab239360 at 1/50 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1\1000 dilution (Green). Confocal image showing the signal translocate to nuclear in HeLa cells treated with IFN-γ (10ng/ml) for 30 mins. Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

      Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

    • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling STAT1 with ab239360 at 1/600 (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 was used as the secondary antibody.

    • STAT1 was immunoprecipitated from 0.35mg RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate with ab239360 at     1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab239360 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

      Lane 1: RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate (10ug)

      Lane 2: ab239360 IP in RAW264.7 whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab239360 in RAW264.7 whole cell lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 75 seconds.

       

    • Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling STAT1 with ab239360 at 1/50 dilution, followed byab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing the signal translocate to nuclear in RAW 264.7 cells treated with IFN-γ (10ng/ml) for 30 mins. Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

      Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

    References

    ab239360 has not yet been referenced specifically in any publications.

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