Overview

  • Product name
    Anti-CD28 antibody [EPR22076]
    See all CD28 primary antibodies
  • Description
    Rabbit monoclonal [EPR22076] to CD28
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IFmore details
    Unsuitable for: Flow Cyt
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human CD28 aa 150 to the C-terminus. The exact sequence is proprietary.
    Database link: P10747

  • Positive control
    • WB: Jurkat and EL4.IL-2 whole cell lysate. IHC-P: Human multiple myeloma and tonsil tissue. Mouse and rat spleen tissue. ICC/IF: Jurkat and EL4.IL-2 cells. IP: Jurkat 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 ab243228 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 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/50.
  • Application notes
    Is unsuitable for Flow Cyt.
  • Target

    Images

    • Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling CD28 with ab243228 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous and cytoplasmic staining on subsets of immune cells of rat spleen is observed. Counterstained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD28 with ab243228 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous and cytoplasmic staining on subsets of immune cells of mouse spleen is observed. Counterstained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Immunohistochemical analysis of paraffin-embedded human multiple myeloma tissue labeling CD28 with ab243228 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous staining on human Multiple Myeloma cells. (PMID: 7522634) is observed. Counterstained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD28 with ab243228 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous and cytoplasmic staining on subsets of immune cells of human tonsil (PMID: 7489738) is observed. Counterstained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • CD28 was immunoprecipitated from 0.35 mg Jurkat (human T cell leukemia T lymphocyte) whole cell lysate with ab243228 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab243228 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.
      Lane 1: Jurkat whole cell lysate 10 µg (Input).
      Lane 2: ab243228 IP in Jurkat whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab243228 in Jurkat whole cell lysate.
      Blocking/Dilution buffer: 5% NFDM/TBST.
      Exposure time: 30 seconds.

      The target band ranging between 37kDa-48kDa is consistent with what has been described in the literature (PMID: 2825196). Minor band below 15kDa is likely to be an isoform.

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized EL4.IL-2 (Mouse lymphoma T lymphocyte) cells labeling CD28 with ab243228 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in EL4.IL-2 cells. The nuclear counter stain is DAPI (blue).
      Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).
      The negative control is the secondary antibody only.

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (human T cell leukemia T lymphocyte) cells labeling CD28 with ab243228 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in Jurkat cells. The nuclear counter stain is DAPI (blue).
      Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).
      The negative control is the secondary antibody only.

    • All lanes : Anti-CD28 antibody [EPR22076] (ab243228) at 1/1000 dilution

      Lane 1 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate
      Lane 2 : EL4.IL-2 (mouse lymphoma T lymphocyte), whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

      Predicted band size: 25 kDa


      Exposure time: 26 seconds


      The target band ranging between 37kDa-48kDa is consistent with what has been described in the literature (PMID: 2825196). Minor band below 15kDa is likely to be an isoform.

      This blot was developed using a higher sensitivity ECL substrate.

      This primary antibody was incubated overnight at 4°C.

    References

    ab243228 has not yet been referenced specifically in any publications.

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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