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使用敲除细胞株进行验证重组RabMAb

重组Anti-STAT1抗体[EPR21057-141] - ChIP Grade (ab234400)

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Western blot - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)
  • Western blot - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)
  • Flow Cytometry (Intracellular) - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)
  • Immunoprecipitation - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)
  • ChIP - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)
  • Flow Cytometry (Intracellular) - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)
  • Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21057-141] to STAT1 - ChIP Grade
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, IP, ChIP
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

概述

  • 产品名称

    Anti-STAT1抗体[EPR21057-141] - ChIP Grade
    参阅全部 STAT1 一抗

  • 描述

    兔单克隆抗体[EPR21057-141] to STAT1 - ChIP Grade

  • 宿主

    Rabbit

  • 经测试应用

    适用于: Flow Cyt (Intra), WB, IHC-P, IP, ChIPmore details

  • 种属反应性

    与反应: Human

  • 免疫原

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • 阳性对照

    • WB: HeLa, HEK-293T and MCF7 cell lysate; Human heart tissue lysate. IHC-P: Human tonsil and kidney tissue; Paried human endometrial cancer and non-tumour endometrium tissue. Flow Cyt (intra): HeLa and MCF7 cells. IP: MCF7 whole cell lysate. ChIP1: HeLa cells.

  • 常规说明

    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.

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab234400于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
Flow Cyt (Intra)
1/500.
WB (4)
1/1000. Predicted molecular weight: 87 kDa.
IHC-P
1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/30.
ChIP
Use at an assay dependent concentration.
说明
Flow Cyt (Intra)
1/500.
WB
1/1000. Predicted molecular weight: 87 kDa.
IHC-P
1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/30.
ChIP
Use at an assay dependent concentration.

靶标

  • 功能

    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.

  • 疾病相关

    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.

  • 序列相似性

    Belongs to the transcription factor STAT family.
    Contains 1 SH2 domain.

  • 翻译后修饰

    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.

  • 细胞定位

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

  • 数据库链接

    • 别名

      • 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

    图片

    • Western blot - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)
      Western blot - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)
      All lanes : Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : STA1 knockout HAP1 whole cell lysate
      Lane 3 : HeLa whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 87 kDa



      Lanes 1 - 3: Merged signal (red and green). Green - ab234400 observed at 87 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab234400 was shown to specifically react with STAT1 in wild-type HAP1 cells as signal was lost in STA1 knockout cells. Wild-type and STA1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab234400 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 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 1/20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)
      Western blot - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)
      All lanes : Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400) at 1/1000 dilution

      Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
      Lane 2 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
      Lane 3 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate at 20 µg
      Lane 4 : Human heart tissue lysate at 10 µg

      Secondary
      Lanes 1-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
      Lane 4 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

      Predicted band size: 87 kDa



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

      Exposure time: Lanes 1-2 & 4: 3 minutes; Lane 3: 15 seconds.

      The doublet observed in some lanes likely represent the α and β isoforms of Stat1 (PMID: 8647800).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)

      Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling STAT1 with ab234400 at 1/5000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining mainly on interfollicular cells of human tonsil (PMID: 25336386; PMID: 25921060) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

      Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Flow Cytometry (Intracellular) - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)
      Flow Cytometry (Intracellular) - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling STAT1 with ab234400 at 1/500 (Red) compared with Rabbit monoclonal IgG (ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. 

       

       

    • Immunoprecipitation - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)
      Immunoprecipitation - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)

      STAT1 was immunoprecipitated from 0.35 mg of MCF7 (human breast adenocarcinoma cell line) whole cell lysate with ab234400 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab234400 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection antibody at 1/5000 dilution.
      Lane 1: MCF7 whole cell lysate 10 μg (input).
      Lane 2: ab234400 IP in MCF7 whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab234400 in MCF7 whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.
      Exposure time: 3 minutes.

      The doublet observed in some lanes likely represent the α and β isoforms of STAT1 (PMID: 8647800).

    • ChIP - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)
      ChIP - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)

      Chromatin was prepared from HeLa (starve overnight) + hIFN-α (serum-starved overnight) (1,000 units/ml,30 min) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25 μg of chromatin, 2 μg of ab234400 (red), and 20 µl of protein A/G sepharose beads slurry (10 µl of sepharose A beads + 10 µl of sepharose G beads). 2 μg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (SYBR approach).
      ChIP results are consistent with the literature (PMID: 16319195).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)

      Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling STAT1 with ab234400 at 1/5000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on glomerulus and some renal tubules of human kidney (PMID: 26678048) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

      Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

       

    • Flow Cytometry (Intracellular) - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)
      Flow Cytometry (Intracellular) - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized MCF7 (human breast adenocarcinoma cell line) cells labeling STAT1 with ab234400 at 1/500 (Red) compared with Rabbit monoclonal IgG (ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. 

       

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)

      Immunohistochemical analysis of paraffin-embedded paired human endometrial cancer (A) and non-tumor endometrium tissue (B) labeling STAT1 with ab234400 at 1/5000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Much higher staining intensity of endometrial cancer (A) than its paired non-tumor endometrium (B) (PMID: 25267067) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

      Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

       

    • Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)
      Anti-STAT1 antibody [EPR21057-141] - ChIP Grade (ab234400)

    实验方案

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    文献 (5)

    发表研究结果有使用 ab234400？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab234400 被引用在 5 文献中.

    • Yang Y  et al. Vitamin D protects glomerular mesangial cells from high glucose-induced injury by repressing JAK/STAT signaling. Int Urol Nephrol 53:1247-1254 (2021). PubMed: 33942213
    • Cai H  et al. LncRNA AIRN influences the proliferation and apoptosis of hepatocellular carcinoma cells by regulating STAT1 ubiquitination. Arch Pharm Res 44:414-426 (2021). PubMed: 33759138
    • Liu W  et al. VAV2 is required for DNA repair and implicated in cancer radiotherapy resistance. Signal Transduct Target Ther 6:322 (2021). PubMed: 34462423
    • Jin Y  et al. Escherichia coli infection activates the production of IFN-a and IFN-ß via the JAK1/STAT1/2 signaling pathway in lung cells. Amino Acids 53:1609-1622 (2021). PubMed: 34524541
    • Qing X  et al. LINC00669 insulates the JAK/STAT suppressor SOCS1 to promote nasopharyngeal cancer cell proliferation and invasion. J Exp Clin Cancer Res 39:166 (2020). PubMed: 32831137

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    1-4 of 4 Abreviews or Q&A

    Western blot abreview for Anti-STAT1 antibody [EPR21057-141] - ChIP Grade

     Poor
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Dog Cell lysate - whole cell (MDCK cells)
    Gel Running Conditions
    Reduced Denaturing (4-15%)
    Loading amount
    25 µg
    Specification
    MDCK cells
    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

    提交于 May 03 2022

    Western blot abreview for Anti-STAT1 antibody [EPR21057-141] - ChIP Grade

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Cell lysate - whole cell (3611-RF cells)
    Gel Running Conditions
    Reduced Denaturing (4-15%)
    Loading amount
    25 µg
    Specification
    3611-RF cells
    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

    提交于 May 03 2022

    Western blot abreview for Anti-STAT1 antibody [EPR21057-141] - ChIP Grade

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (MEFs)
    Gel Running Conditions
    Reduced Denaturing (4-15%)
    Loading amount
    25 µg
    Specification
    MEFs
    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

    提交于 May 03 2022

    Western blot abreview for Anti-STAT1 antibody [EPR21057-141] - ChIP Grade

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa cells)
    Gel Running Conditions
    Reduced Denaturing (4-15%)
    Loading amount
    25 µg
    Specification
    HeLa cells
    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

    提交于 May 03 2022

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