Overview

  • Product name

    Anti-CD134 antibody [EPR22229-5]
    See all CD134 primary antibodies
  • Description

    Rabbit monoclonal [EPR22229-5] to CD134
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Rat
  • Immunogen

    Recombinant fragment within Mouse CD134 aa 1-250. The exact sequence is proprietary.
    Database link: P47741

  • Positive control

    • WB: Mouse and rat splenocytes treated with 2.5ug/ml concanavalin A for 72h lysates. IHC-P: Mouse and rat spleen tissues. Flow Cyt: Rat and mouse primary splenocytes treated with 2.5µg/ml concanavalin A for 72h. IP: Rat splenocytes treated with 2.5µg/ml concanavalin A for 72h lysate.
  • 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 ab229021 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. Detects a band of approximately 50 kDa (predicted molecular weight: 29 kDa).
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt 1/500.
IP 1/30.
  • Application notes
    Is unsuitable for ICC/IF or IHC-Fr.
  • Target

    • Function

      Receptor for TNFSF4/OX40L/GP34.
    • Sequence similarities

      Contains 4 TNFR-Cys repeats.
    • Cellular localization

      Membrane.
    • Information by UniProt
    • Database links

    • Alternative names

      • ACT35 antibody
      • ACT35 antigen antibody
      • CD134 antibody
      • CD134 antigen antibody
      • Lymphoid activation antigene ACT35 antibody
      • OX40 antibody
      • OX40 antigen antibody
      • OX40 cell surface antigen antibody
      • OX40 homologue antibody
      • OX40L receptor antibody
      • TAX transcriptionally-activated glycoprotein 1 receptor antibody
      • TNFRSF4 antibody
      • TNR4_HUMAN antibody
      • Tumor necrosis factor receptor superfamily member 4 antibody
      • TXGP1L antibody
      see all

    Images

    • Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling CD134 using ab229021 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on T cells in rat spleen (PMID: 15259024; PMID: 10233901). Counterstained 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10mins.

      The section was incubated with ab229021 for 10 mins at 37℃. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument.

    • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD134 using ab229021 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on T cells in periarterial lymphatic sheath of mouse spleen (PMID: 15259024; PMID: 10233901). Counterstained 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10mins.

      The section was incubated with ab229021 for 10 mins at 37℃. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument.

    • CD134 was immunoprecipitated from 0.35 mg rat splenocytes treated with 2.5μg/ml concanavalin A for 72h lysate using ab229021 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab229021 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at a 1/5000 dilution.

      Lane 1: Rat splenocytes treated with 2.5μg/ml concanavalin A for 72h lysate 10 μg (input)
      Lane 2: ab229021 IP in Rat splenocytes treated with 2.5μg/ml concanavalin A for 72h lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab229021 in Rat splenocytes treated with 2.5μg/ml concanavalin A for 72h lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.
      Exposure time: 20 seconds.

    • Flow cytometric analysis of mouse primary splenocytes treated with 2.5μg/ml concanavalin A for 72h labeling CD134 with ab229021 at 1/500 dilution (Right) compared to cells stained with Alexa Fluor® 647-conjugated CD3 and rabbit IgG (Left). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at 1/2000 dilution.

      Data shown were gated on viable cells.

    • Flow cytometric analysis of rat primary splenocytes treated with 2.5μg/ml concanavalin A for 72h labeling CD134 with ab229021 at 1/500 dilution (Right) compared to cells stained with Alexa Fluor® 647-conjugated CD3 and rabbit IgG (Left). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at 1/2000 dilution.

      Data shown were gated on viable cells.

    • All lanes : Anti-CD134 antibody [EPR22229-5] (ab229021) at 1/1000 dilution

      Lane 1 : Untreated mouse splenocytes lysate
      Lane 2 : Mouse splenocytes treated with 2.5ug/ml concanavalin A for 72h lysate
      Lane 3 : Untreated rat splenocytes lysate
      Lane 4 : Rat splenocytes treated with 2.5ug/ml concanavalin A for 72h 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: 29 kDa
      Observed band size: 50 kDa
      why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/Diluting buffer and concentration: 5% NFDM/TBST

      CD134 is a glycosylated protein and its expression is increased after concanavalin A treatment. PMID 23897980; PMID 17609274

    References

    ab229021 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (spleen)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
    Permeabilization
    No
    Specification
    spleen
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: RT°C
    Fixative
    10% NBF

    Abcam user community

    Verified customer

    Submitted Mar 15 2019

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

    Sign up