RecombinantRabMAb

Recombinant Anti-CD28 antibody [EPR22311-17] (ab242124)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD28 antibody [EPR22311-17] (ab242124)
  • Immunoprecipitation - Anti-CD28 antibody [EPR22311-17] (ab242124)
  • Western blot - Anti-CD28 antibody [EPR22311-17] (ab242124)
  • Anti-CD28 antibody [EPR22311-17] (ab242124)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22311-17] to CD28
  • Suitable for: WB, IHC-P, IP
  • Reacts with: Mouse

Overview

  • Product name

    Anti-CD28 antibody [EPR22311-17]
    See all CD28 primary antibodies

  • Description

    Rabbit monoclonal [EPR22311-17] to CD28

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Mouse

  • Immunogen

    Recombinant fragment within Mouse CD28 aa 1-150. The exact sequence is proprietary.
    Database link: P31041

  • Positive control

    • WB: Mouse thymus tissue lysate; EL4.IL-2 whole cell lysate. IHC-P: Mouse spleen tissue. IP: EL4.IL-2 whole cell 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.

    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 ab242124 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. Predicted molecular weight: 25 kDa.
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Perform heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10 mins.
IP 1/30.
  • Application notes
    Is unsuitable for Flow Cyt or ICC/IF.

    • Target

    • Function

      Involved in T-cell activation, the induction of cell proliferation and cytokine production and promotion of T-cell survival.

    • Tissue specificity

      Expressed in T-cells and plasma cells, but not in less mature B-cells.

    • Sequence similarities

      Contains 1 Ig-like V-type (immunoglobulin-like) domain.

    • Cellular localization

      Membrane.
    • Information by UniProt

    • Database links

      • Alternative names

        • CD 28 antibody
        • CD28 antibody
        • CD28 antigen antibody
        • CD28 molecule antibody
        • CD28_HUMAN antibody
        • MGC138290 antibody
        • T cell antigen CD28 antibody
        • T cell specific surface glycoprotein antibody
        • T cell specific surface glycoprotein CD28 antibody
        • T-cell-specific surface glycoprotein CD28 antibody
        • TP44 antibody
        see all

      Images

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD28 antibody [EPR22311-17] (ab242124)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD28 antibody [EPR22311-17] (ab242124)

        Immunohistochemical analysis of paraffin-embedded mouse spleen tissue stained for CD28 using ab242124 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining in periarterial lymphatic sheath of mouse spleen (PMID: 7489738). 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.

        The section was incubated with ab242124 for 30 mins at 37℃. Immunostaining was performed on a Leica Biosystems BOND® RX instrument.

        Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

      • Immunoprecipitation - Anti-CD28 antibody [EPR22311-17] (ab242124)
        Immunoprecipitation - Anti-CD28 antibody [EPR22311-17] (ab242124)

        CD28 was immunoprecipitated from 0.35 mg EL4.IL-2 (mouse lymphoma T lymphocyte) whole cell lysate with ab242124 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab242124 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.

        Lane 1: EL4.IL-2 whole cell lysate 10 µg (input).
        Lane 2: ab242124 IP in EL4.IL-2 whole cell lysate.
        Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab242124 in EL4.IL-2 whole cell lysate.

        Blocking and dilution buffer and concentration: 5% NFDM/TBST.
        Exposure time: 15 seconds.

         

      • Western blot - Anti-CD28 antibody [EPR22311-17] (ab242124)
        Western blot - Anti-CD28 antibody [EPR22311-17] (ab242124)
        All lanes : Anti-CD28 antibody [EPR22311-17] (ab242124) at 1/1000 dilution

        Lane 1 : Mouse thymus tissue lysate
        Lane 2 : EL4.IL-2 (mouse lymphoma t lymphocyte) whole cell 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: 25 kDa
        Observed band size: 40-50,80 kDa
        why is the actual band size different from the predicted?


        Exposure time: 70 seconds


        Blocking/Dilution buffer: 5% NFDM/TBST.

        Bands between 40-50 kDa are likely to be glycosylated forms (PMID: 7512728), whereas the 80 kDa band represents the dimer (PMID: 2157764).

      • Anti-CD28 antibody [EPR22311-17] (ab242124)
        Anti-CD28 antibody [EPR22311-17] (ab242124)

      Protocols

      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

      References (0)

      Publishing research using ab242124? Please let us know so that we can cite the reference in this datasheet.

      ab242124 has not yet been referenced specifically in any publications.

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