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CD28 Antibody | D665

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  • Mouse anti Mouse CD28:Low Endotoxin
  • Mouse anti Mouse CD28:Alexa Fluor® 488

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  • Product Type
    Monoclonal Antibody
  • Clone
    D665
  • Isotype
    IgG1
2 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    MCA2587ELFNdatasheet pdfdatasheet pdf0.5 mg
    MCA2587EL
    MCA2587A488Fdatasheet pdfdatasheet pdf100 Tests/1ml
    MCA2587A488
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
    -
    • Mouse anti Mouse CD28 antibody, clone D665 recognizes murine CD28, also known as T-cell-specific surface glycoprotein CD28, a 218 amino acid ~90 kDa single pass type 1 transmembrane glycoprotein containing a single Ig-like v-type domain. CD28 is a member of the immunoglobulin superfamily and is structurally related to CD152 (CTLA-4). CD28 is a major costimulatory molecule, expressed by most T cells including thymocyte subsets and peripheral T-cells but is absent on a subset of CD8+ve cells. Engagement of CD28 by its natural ligand, CD80 or CD86, provides a costimulatory signal which promotes T-cell effector function and T-cell-dependent antibody production.

      Mouse anti mouse CD28 antibody clone D665 is reported to act as a CD28 superagonist antibody, which induces antigen-independent T cell proliferation (Dennehy et al. 2006). Mouse anti mouse CD28 antibody, clone D665 binds to an epitope in the C"D loop of murine CD28, similar to the recognized epitope for other superagonist antibodies directed against human and rat CD28 targets (Lühder et al. 2003).

      Activation of Treg cells through engagement of CD28 by mouse anti mouse CD28 antibody, clone D665 leads to improved survival and healing in mice folowing myocardial infarction by modulating monocyte/ macrophage differentiation (Weirather et al. 2014). In mice it appears that Treg cells offer sufficient regulatory control following superagonist engagement of anti CD28 antibody to dampen the proinflammatory cytokine release, seen in the absence of Treg cells (Gogishvili et al. 2009) and in the cytokine release syndrome observed in participants of the phase 1 CD28 superagonist antibody TGN1412 clinical trial (Suntharalingam et al. 2006).
    • Intended Use
    • Target Species
      Mouse
    • Product Form
      Purified IgG - liquid
      Purified IgG conjugated to Alexa Fluor® 488 - liquid
    • Reconstitution
    • Preparation
      Purified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant
      Purified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant
    • Preservative Stabilisers
      None present.
      0.09%Sodium Azide (NaN3)
      1%Bovine Serum Albumin
    • Immunogen
      Alternative immunisation with A20 cells expressing mCD28 and recombinant mCD28Ig.
    • Purity
    • Approx. Protein Concentrations
      IgG concentration 1.0mg/ml
      IgG concentration 0.05mg/ml
    • Reagents In The Kit
    • Preparing The Antibody
    • Endotoxin Level
      <0.01Eu/ug
    • Test Principle
    • Buffer Solution
      Phosphate buffered saline
      Phosphate buffered saline
    • Fusion Partners
      Spleen cells from immunised CD28-/-knockout mice were fused with cells of the X63Ag8.653 myeloma cell line.
    • Storage
      Store at -20oC only.
      Storage in frost-free freezers is not recommended.
      This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
      Store at +4oC or at -20oC if preferred.
      Storage in frost-free freezers is not recommended.
      This product should be stored undiluted. This product is photosensitive and should be protected from light.
      Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
    • Shelf Life
      18 months from date of despatch.
      18 months from date of despatch.
    • GO Terms
      GO:0045944
      positive regulation of transcription from RNA polymerase II promoter
      Related reagents
      GO:0002863
      positive regulation of inflammatory response to antigenic stimulus
      Related reagents
      GO:0045086
      positive regulation of interleukin-2 biosynthetic process
      Related reagents
      GO:0045840
      positive regulation of mitosis
      Related reagents
      GO:0016021
      integral to membrane
      Related reagents
      GO:0046641
      positive regulation of alpha-beta T cell proliferation
      Related reagents
      GO:0009897
      external side of plasma membrane
      Related reagents
      GO:0031295
      T cell costimulation
      Related reagents
      GO:0008624
      induction of apoptosis by extracellular signals
      Related reagents
      GO:0048304
      positive regulation of isotype switching to IgG isotypes
      Related reagents
      GO:0006955
      immune response
      Related reagents
      GO:0045060
      negative thymic T cell selection
      Related reagents
    • UniProt
    • Entrez Gene
    • Acknowledgements
      This product is provided under an intellectual property licence from Life Technologies Corporation. The transfer of this product is contingent on the buyer using the purchase product solely in research, excluding contract research or any fee for service research, and the buyer must not sell or otherwise transfer this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; (c) manufacturing or quality assurance or quality control, or (d) resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad CA 92008 USA or outlicensing@thermofisher.com
    • Regulatory
      For research purposes only
    • This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.

    • Application NameYesNoMin DilutionMax Dilution
      Functional Assays1-10ug/ml
    • Application NameYesNoMin DilutionMax Dilution
      Flow CytometryNeat1/10

    • Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
    • Technical Advice
    • Technical Advice
    • Recommended Protocol
    • Recommended Protocol
    • ELISA
    • ELISA
    • Immunohistology
    • Immunohistology
    • Histology Positive Control Tissue
    • Histology Positive Control Tissue
    • Immunofluorescence
    • Immunofluorescence
    • Western Blotting
    • Western Blotting
    • Instructions For Use
    • Instructions For Use

    Additional CD28 Antibody Formats

    Formats Clone Applications Sizes available
    CD28 Antibody : Low Endotoxin D665 FN 0.5 mg
    CD28 Antibody : Alexa Fluor® 488 D665 F 100 Tests/1ml
    • Copyright © 2017 Bio-Rad Antibodies (formerly AbD Serotec)

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      Recommended Negative Isotype Control

        Useful Reagents

          DescriptionProduct CodePack SizeApplicationsList PriceQuantity
          Mouse Seroblock FcRBUF041A0.1 mgF
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          Mouse Seroblock FcRBUF041B0.5 mgF
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          Recommended Positive Controls

            Histology Controls

              References

              1. Dennehy, K.M. et al. (2006) Cutting edge: monovalency of CD28 maintains the antigen dependence of T cell costimulatory responses.
              J Immunol. 176 (10): 5725-9.
              2. Vaeth, M. et al, (2012) Dependence on nuclear factor of activated T-cells (NFAT) levels discriminates conventional T cells from Foxp3+ regulatory T cells.
              Proc Natl Acad Sci U S A. 109: 16258-63.
              3. Gogishvili, T. et al. (2009) Rapid regulatory T-cell response prevents cytokine storm in CD28 superagonist treated mice.
              PLoS One. 4: e4643.
              4. Zhang, T. et al. (2011) Selective CD28 blockade attenuates acute and chronic rejection of murine cardiac allografts in a CTLA-4-dependent manner.
              Am J Transplant. 11: 1599-609.
              5. Weirather, J. et al. (2014) Foxp3+ CD4+ T cells improve healing after myocardial infarction by modulating monocyte/macrophage differentiation.
              Circ Res. 115 (1): 55-67.

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