Recombinant
RabMAb

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

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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

    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 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).

      References

      ab242124 has not yet been referenced specifically in any publications.

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

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