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  1. Link

    stat1-alpha-antibody-epyr2154-ab92506.pdf

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Epigenetics and Nuclear Signaling Transcription Domain Families HLH / Leucine Zipper Leucine Zipper
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Validated using a knockout cell lineRecombinantRabMAb

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

  • Datasheet
Reviews (2) Submit a question References (15)

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

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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

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

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

    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.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPYR2154
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • HLH / Leucine Zipper
    • Leucine Zipper
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • STATs
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • STATs
    • Cancer
    • Signal transduction
    • Nuclear signaling
    • STAT family

Associated products

  • Alternative Versions

    • Anti-STAT1 alpha antibody [EPYR2154] - BSA and Azide free (ab168333)
    • HRP Anti-STAT1 alpha antibody [EPYR2154] (ab193891)
  • Assay kits

    • STAT (STAT1alpha, STAT3, STAT5A, STAT5B) Transcription Factor Assay Kit (Colorimetric) (ab207228)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • NIH/3T3 nuclear extract lysate (ab14874)
  • Recombinant Protein

    • Recombinant Human STAT1 alpha protein - GST tagged (ab140412)

Applications

The Abpromise guarantee

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 (2)
1/1000 - 1/2000. Predicted molecular weight: 87 kDa.
IP
1/20 - 1/50.
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.
  • Target information above from: UniProt accession P42224 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 6772 Human
    • Entrez Gene: 20846 Mouse
    • Omim: 600555 Human
    • SwissProt: P42224 Human
    • SwissProt: P42225 Mouse
    • Unigene: 642990 Human
    • Unigene: 727504 Human
    • 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)
      All lanes : Anti-STAT1 alpha antibody [EPYR2154] (ab92506) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : STAT1 knockout HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 87 kDa
      Observed band size: 85 kDa why is the actual band size different from the predicted?



      Lanes 1 - 2: Merged signal (red and green). Green - ab92506 observed at 85 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

      ab92506 was shown to react with STAT1 alpha in wild-type HeLa cells in Western blot with loss of signal observed in STAT1 knockout cell line ab255346 (STAT1 knockout cell lysate ab263837). Wild-type HeLa and STAT1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab92506 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 h at room temperature before imaging.

    • 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)

    Protocols

    • Western blot protocols
    • Flow cytometry protocols
    • Immunoprecipitation protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (15)

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

    ab92506 has been referenced in 15 publications.

    • Hu C  et al. Inhibition of Long Noncoding RNA SNHG15 Ameliorates Hypoxia/Ischemia-Induced Neuronal Damage by Regulating miR-302a-3p/STAT1/NF-?B Axis. Yonsei Med J 62:325-337 (2021). PubMed: 33779086
    • Su Q  et al. IFN-? induces apoptosis in human melanocytes by activating the JAK1/STAT1 signaling pathway. Mol Med Rep 22:3111-3116 (2020). PubMed: 32945463
    • 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
    • Tian J  et al. Feline calicivirus strain 2280 p30 antagonizes type I interferon-mediated antiviral innate immunity through directly degrading IFNAR1 mRNA. PLoS Pathog 16:e1008944 (2020). PubMed: 33075108
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-2 of 2 Abreviews or Q&A

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

    Average
    Abreviews
    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
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Mar 13 2019

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

    Excellent
    Abreviews
    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
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Aug 03 2015

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