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Anti-IRF3抗体[IRF35I218] (ab50772)

Reviews (3)Q&A (4)References (11)
Western blot - Anti-IRF3 antibody [IRF35I218] (ab50772)
  • Western blot - Anti-IRF3 antibody [IRF35I218] (ab50772)
  • Flow Cytometry - Anti-IRF3 antibody [IRF35I218] (ab50772)
  • Immunocytochemistry/ Immunofluorescence - Anti-IRF3 antibody [IRF35I218] (ab50772)
  • Western blot - Anti-IRF3 antibody [IRF35I218] (ab50772)

Key features and details

  • Mouse monoclonal [IRF35I218] to IRF3
  • Suitable for: ICC/IF, WB, Flow Cyt
  • Reacts with: Human
  • Isotype: IgG1

概述

  • 产品名称

    Anti-IRF3抗体[IRF35I218]
    参阅全部 IRF3 一抗

  • 描述

    小鼠单克隆抗体[IRF35I218] to IRF3

  • 宿主

    Mouse

  • 经测试应用

    适用于: ICC/IF, WB, Flow Cytmore details

  • 种属反应性

    与反应: Human

  • 免疫原

    Recombinant fragment: VAWVNKSRTR FRIPWKHGLR QDAQQEDFGI FQAWAEATGA YVPGRDKPDL PTWKRNFRSA LNRKEGLRLA EDRSKD, corresponding to amino acids 24-150 of Human IRF3

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • 阳性对照

    • HEK293 whole cell lysate RIPA lysate of HeLa cells Hela cells

  • 常规说明

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab50772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
ICC/IF
1/10.
WB (3)
1/50 - 1/100. Detects a band of approximately 55 kDa (predicted molecular weight: 47 kDa).
Flow Cyt
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
说明
ICC/IF
1/10.
WB
1/50 - 1/100. Detects a band of approximately 55 kDa (predicted molecular weight: 47 kDa).
Flow Cyt
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

靶标

  • 功能

    Mediates interferon-stimulated response element (ISRE) promoter activation. Functions as a molecular switch for antiviral activity. DsRNA generated during the course of an viral infection leads to IRF3 phosphorylation on the C-terminal serine/threonine cluster. This induces a conformational change, leading to its dimerization, nuclear localization and association with CREB binding protein (CREBBP) to form dsRNA-activated factor 1 (DRAF1), a complex which activates the transcription of genes under the control of ISRE. The complex binds to the IE and PRDIII regions on the IFN-alpha and IFN-beta promoters respectively. IRF-3 does not have any transcription activation domains.

  • 组织特异性

    Expressed constitutively in a variety of tissues.

  • 序列相似性

    Belongs to the IRF family.
    Contains 1 IRF tryptophan pentad repeat DNA-binding domain.

  • 翻译后修饰

    Constitutively phosphorylated on many serines residues. C-terminal serine/threonine cluster is phosphorylated in response of induction by IKBKE and TBK1. Ser-385 and Ser-386 may be specifically phosphorylated in response to induction. An alternate model propose that the five serine/threonine residues between 396 and 405 are phosphorylated in response to a viral infection. Phosphorylation, and subsequent activation of IRF3 is inhibited by vaccinia virus protein E3.
    Ubiquitinated; ubiquitination involves RBCK1 leading to proteasomal degradation. Polyubiquitinated; ubiquitination involves TRIM21 leading to proteasomal degradation.
    ISGylated by HERC5 resulting in sustained IRF3 activation and in the inhibition of IRF3 ubiquitination by disrupting PIN1 binding. The phosphorylation state of IRF3 does not alter ISGylation.

  • 细胞定位

    Cytoplasm. Nucleus. Shuttles between cytoplasmic and nuclear compartments, with export being the prevailing effect. When activated, IRF3 interaction with CREBBP prevents its export to the cytoplasm.
  • Information by UniProt

  • 数据库链接

    • 别名

      • IIAE7 antibody
      • Interferon regulatory factor 3 antibody
      • IRF 3 antibody
      • IRF-3 antibody
      • IRF3 antibody
      • IRF3_HUMAN antibody
      • MGC94729 antibody
      see all

    图片

    • Western blot - Anti-IRF3 antibody [IRF35I218] (ab50772)
      Western blot - Anti-IRF3 antibody [IRF35I218] (ab50772)This image is courtesy of an anonymous Abreview
      Anti-IRF3 antibody [IRF35I218] (ab50772) at 1/2000 dilution + Human THP1 lysate at 25 µg

      Secondary
      HRP Goat Anti Mouse Secondary Antibody at 1/10000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 47 kDa
      Observed band size: 45 kDa why is the actual band size different from the predicted?


      Exposure time: 1 minute

      See Abreview

    • Western blot - Anti-IRF3 antibody [IRF35I218] (ab50772)
      Western blot - Anti-IRF3 antibody [IRF35I218] (ab50772)
      Anti-IRF3 antibody [IRF35I218] (ab50772) at 1/50 dilution + HT-1080 whole cell lysate at 25 µg

      Secondary
      Anti mouse IgG at 1/2500 dilution

      Predicted band size: 47 kDa
      Observed band size: 55 kDa why is the actual band size different from the predicted?

    • Flow Cytometry - Anti-IRF3 antibody [IRF35I218] (ab50772)
      Flow Cytometry - Anti-IRF3 antibody [IRF35I218] (ab50772)
      Overlay histogram showing HeLa cells stained with ab50772 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab50772, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
    • Immunocytochemistry/ Immunofluorescence - Anti-IRF3 antibody [IRF35I218] (ab50772)
      Immunocytochemistry/ Immunofluorescence - Anti-IRF3 antibody [IRF35I218] (ab50772)
      Immunofluorescent staining of HeLa cells with a 1/10 dilution of ab50772 as primary antibody and AlexaFluor®488 conjugated Goat Anti-mouse IgG diluted 1/200 as the secondary antibody
    • Western blot - Anti-IRF3 antibody [IRF35I218] (ab50772)
      Western blot - Anti-IRF3 antibody [IRF35I218] (ab50772)
      Anti-IRF3 antibody [IRF35I218] (ab50772) at 1/100 dilution + Ripa lysate of Hela cells at 500 µg

      Secondary
      Anti-mouse IgG at 1/1000 dilution

      Predicted band size: 47 kDa
      Observed band size: 55 kDa why is the actual band size different from the predicted?



      IRF3 antibody [IRF35I218] (ab50772) at 1/100 dilution + Ripa lysate of Hela cells at 500 µg Secondary Anti-mouse IgG at 1/1000 dilution

    文献 (11)

    发表研究结果有使用 ab50772？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab50772 被引用在 11 文献中.

    • Wang J  et al. Human OTUD6B positively regulates type I IFN antiviral innate immune responses by deubiquitinating and stabilizing IRF3. mBio 14:e0033223 (2023). PubMed: 37650650
    • Lu D  et al. Activation of STING by targeting a pocket in the transmembrane domain. Nature 604:557-562 (2022). PubMed: 35388221
    • Uhlorn BL  et al. Attenuation of cGAS/STING activity during mitosis. Life Sci Alliance 3:N/A (2020). PubMed: 32661021
    • Uhlorn BL  et al. Vesicular trafficking permits evasion of cGAS/STING surveillance during initial human papillomavirus infection. PLoS Pathog 16:e1009028 (2020). PubMed: 33253291
    • Yi L  et al. Interferon regulatory factor 3 mediates Poly(I:C)-induced innate immune response and apoptosis in non-small cell lung cancer. Int J Oncol 52:1623-1632 (2018). PubMed: 29512705
    View all Publications for this product

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价 提交问题

    1-7 of 7 Abreviews or Q&A

    Western blot abreview for Anti-IRF3 antibody [IRF35I218]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (THP1)
    Gel Running Conditions
    Reduced Denaturing (10)
    Loading amount
    25 µg
    Specification
    THP1
    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

    提交于 Jul 19 2018

    Western blot abreview for Anti-IRF3 antibody [IRF35I218]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Human Dermal Microvascular Endothelial Cells (Prim)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    30 µg
    Specification
    Human Dermal Microvascular Endothelial Cells (Prim
    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

    提交于 Mar 14 2017

    Western blot abreview for Anti-IRF3 antibody [IRF35I218]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK293T and hela)
    Loading amount
    25 µg
    Specification
    HEK293T and hela
    Gel Running Conditions
    Reduced Denaturing
    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

    提交于 Nov 15 2007

    Question

    hi, i am looking for an IRF-3 specific antibody that can detect both the cytoplasmic and nuclear form (i.e. to see a shift from cytoplasm to the nucleus). most of your products show nice cytoplasmic staining. can you direct me to one that you may have tested under activating conditions? (eg. polyIC or dsRNA) thanks

    Read More
    Answer

    Unfortunately, when we test our IRF-3 antibodies we do not specifically test them in activating conditions so we have no results to share with you. However, several of our antibodies have been used in publications under such conditions and so I believe would be suitable for your experiments.


    Anti-IRF3 antibody (ab46194) was used in the publication: Boxer EL et al. The rinderpest virus non-structural C protein blocks the induction of type 1 interferon. Virology 385:134-42 (2009) under activating conditions.


    Anti-IRF3 antibody [IRF35I218] (ab50772) was used in Chen GY et al. Baculovirus transduction of mesenchymal stem cells triggers the toll-like receptor 3 pathway. J Virol 83:10548-56 (2009) under activating conditions.

    Read More

    Question

    Western blot, no band
    ab21680 1/1000
    O/N 4ºC, 1h RT
    5% milk
    FOCR with ab50772 requested.
    (ab76493 is working fine)

    Read More
    Answer

    Merci de nous avoir contactés.

    Nous sommes désolés que le produit que vous avez reçu ne fonctionne pas comme attendu.

    Comme convenu j'ai mis en place l'envoi d'une unité de ab50772 en remplacement du ab21680. Le numéro de commande de remplacement gratuit de ce produit est ********.

    Vous recevrez prochainement un mail de confirmation comprenant les détails d'expédition.

    N'hésitez pas à nous contacter lors d'une prochaine occasion.

    Read More

    Question



    There are no attached file . Please send me the protocol file.

    Read More
    Answer

    Apologies for the attachment not coming through in the last email. I have attached file again.

    Please let me know if there is anything else I can help you with.

    Read More

    Question

    What protocol was used to generate the IF image on the webpage? Was there an activation step?

    Read More
    Answer

    Thank you for contacting Abcam.

    The protocol used to generate the IF image that we talked about on the datasheet for ab50772 is attached, there was no activation step performed before the staining:

    Read More

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