Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-STAT1 alpha antibody [EPYR2154] (ab92506)

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Western blot - Anti-STAT1 alpha antibody [EPYR2154] (ab92506)
  • Immunoprecipitation - Anti-STAT1 alpha antibody [EPYR2154] (ab92506)
  • Western blot - Anti-STAT1 alpha antibody [EPYR2154] (ab92506)
  • Western blot - Anti-STAT1 alpha antibody [EPYR2154] (ab92506)
  • Western blot - Anti-STAT1 alpha antibody [EPYR2154] (ab92506)
  • Anti-STAT1 alpha antibody [EPYR2154] (ab92506)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPYR2154] to STAT1 alpha
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-STAT1 alpha antibody [EPYR2154]
    See all STAT1 alpha primary antibodies

  • Description

    Rabbit monoclonal [EPYR2154] to STAT1 alpha

  • Host species

    Rabbit

  • Specificity

    Based on Blast results using the immunogen sequence, this antibody should recognise the alpha but not the beta form (only 30% homology) of Stat1.

  • Tested applications

    Suitable for: WB, IPmore details
    Unsuitable for: Flow Cyt

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    corresponding to Human STAT1 alpha aa 700 to the C-terminus.
    Database link: P42224

  • Positive control

    • HeLa, HEK293, NIH-3T3 and A431 cell lysates, mouse brain tissue lysate

  • General notes

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab92506 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
WB 1/1000 - 1/2000. Predicted molecular weight: 87 kDa.
IP 1/20 - 1/50.
  • Application notes
    Is unsuitable for Flow Cyt.

    • 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

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

      Images

      • Western blot - Anti-STAT1 alpha antibody [EPYR2154] (ab92506)
        Western blot - Anti-STAT1 alpha antibody [EPYR2154] (ab92506)

        Lane 1: Wild type HAP1 whole cell lysate (20 µg)
        Lane 2: STAT1 alpha knockout HAP1 whole cell lysate (20 µg)
        Lane 3: HeLa whole cell lysate (20 µg)
        Lane 4: A431 whole cell lysate (20 µg)

        Lanes 1 - 4: Merged signal (red and green). Green - ab92506 (unpurified) observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.

        ab92506 was shown to specifically react with STAT1 alpha when STAT1 alpha knockout samples were used. Wild-type and STAT1 alpha knockout samples were subjected to SDS-PAGE. Ab92506 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

      • Immunoprecipitation - Anti-STAT1 alpha antibody [EPYR2154] (ab92506)
        Immunoprecipitation - Anti-STAT1 alpha antibody [EPYR2154] (ab92506)

        ab92506 (purified) at 1/20 dilution (0.5ug) immunoprecipitating STAT1 alpha in A431 whole cell lysate. A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate 10ug
        Lane 2 (+): ab92506 & A431 whole cell lysate
        Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab92506 in A431 whole cell lysate
        For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/2000 dilution.
        Blocking and diluting buffer: 5% NFDM/TBST.

      • Western blot - Anti-STAT1 alpha antibody [EPYR2154] (ab92506)
        Western blot - Anti-STAT1 alpha antibody [EPYR2154] (ab92506)
        Lane 1 : Anti-STAT1 alpha antibody [EPYR2154] (ab92506) at 1/1000 dilution (Purified)
        Lanes 2-3 : Anti-STAT1 alpha antibody [EPYR2154] (ab92506) at 1/1000 dilution

        Lane 1 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate
        Lane 2 : Mouse brain lysate
        Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates

        Lysates/proteins at 20 µg per lane.

        Secondary
        All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

        Predicted band size: 87 kDa
        Observed band size: 91 kDa
        why is the actual band size different from the predicted?

      • Western blot - Anti-STAT1 alpha antibody [EPYR2154] (ab92506)
        Western blot - Anti-STAT1 alpha antibody [EPYR2154] (ab92506)
        All lanes : Anti-STAT1 alpha antibody [EPYR2154] (ab92506) at 1/2000 dilution (unpurified)

        Lane 1 : Untreated HeLa (human cervix adenocarcinoma) membrane
        Lane 2 : HeLa (human cervix adenocarcinoma) membrane treated with Alkaline Phosphatase

        Lysates/proteins at 10 µg per lane.

        Secondary
        All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1500 dilution

        Developed using the ECL technique.

        Predicted band size: 87 kDa
        Observed band size: 91 kDa why is the actual band size different from the predicted?


        Exposure time: 3 minutes


        Phospho STAT1 protein is revealed by ab109461, anti-STAT1 (phospho S727) antibody. ab92506 is used as pan control which detects total STAT1.

        Blocking and dilution buffer: 5% NFDM/TBST.

      • Western blot - Anti-STAT1 alpha antibody [EPYR2154] (ab92506)
        Western blot - Anti-STAT1 alpha antibody [EPYR2154] (ab92506)
        All lanes : Anti-STAT1 alpha antibody [EPYR2154] (ab92506) at 1/1000 dilution (unpurified)

        Lane 1 : HeLa cell lysate
        Lane 2 : 293 cell lysate
        Lane 3 : NIH-3T3 cell lysate
        Lane 4 : A431 cell lysate

        Lysates/proteins at 10 µg per lane.

        Secondary
        All lanes : HRP labelled goat anti-rabbit IgG at 1/2000 dilution

        Predicted band size: 87 kDa
        Observed band size: 91 kDa why is the actual band size different from the predicted?

      • Anti-STAT1 alpha antibody [EPYR2154] (ab92506)
        Anti-STAT1 alpha antibody [EPYR2154] (ab92506)

      References (11)

      Publishing research using ab92506? Please let us know so that we can cite the reference in this datasheet.

      ab92506 has been referenced in 11 publications.

      • Dan H  et al. RACK1 promotes cancer progression by increasing the M2/M1 macrophage ratio via the NF-?B pathway in oral squamous cell carcinoma. Mol Oncol 14:795-807 (2020). PubMed: 31997535
      • Ma C  et al. piRNA-63076 contributes to pulmonary arterial smooth muscle cell proliferation through acyl-CoA dehydrogenase. J Cell Mol Med 24:5260-5273 (2020). PubMed: 32227582
      • Zhang J  et al. miR-26a Inhibits Feline Herpesvirus 1 Replication by Targeting SOCS5 and Promoting Type I Interferon Signaling. Viruses 12:N/A (2019). PubMed: 31861450
      • Xing Y  et al. Late stages of T cell maturation in the thymus involve NF-?B and tonic type I interferon signaling. Nat Immunol 17:565-73 (2016). PubMed: 27043411
      • Akhrymuk I  et al. Both RIG-I and MDA5 detect alphavirus replication in concentration-dependent mode. Virology 487:230-41 (2016). WB ; Mouse . PubMed: 26550947
      View all Publications for this product

      Customer reviews and Q&As

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      1-2 of 2 Abreviews or Q&A

      Western blot abreview for Anti-STAT1 alpha antibody [EPYR2154]

       Average
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Human Cell lysate - whole cell (ARPE-19)
      Gel Running Conditions
      Reduced Denaturing (4-12% Bis-Tris Gel)
      Loading amount
      10 µg
      Treatment
      125 ug/ml CSE for 24 hr
      Specification
      ARPE-19
      Blocking step
      Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
      Read More

      Abcam user community

      Verified customer

      Submitted Mar 13 2019

      Western blot abreview for Anti-STAT1 antibody [EPYR2154]

       Excellent
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Human Cell lysate - whole cell (HCT116)
      Gel Running Conditions
      Reduced Denaturing
      Loading amount
      10 µg
      Specification
      HCT116
      Blocking step
      Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
      Read More

      Abcam user community

      Verified customer

      Submitted Aug 03 2015

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