Human CD28 Antibody

(4 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human CD28 in direct ELISAs and Western blots. In direct ELISAs, approximately 20% cross-reactivity with recombinant mouse CD28 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    S. frugiperda insect ovarian cell line Sf 21-derived recombinant human CD28
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    Recombinant Human CD28 Fc Chimera (Catalog # 342-CD)
  • Flow Cytometry
    2.5 µg/106 cells
    Human whole blood CD3+ T cells
  • Agonist Activity
    0.3-0.6 µg/mL
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Human CD28 Antibody Enhances IL-2 Secretion in Jurkat Cells. Human CD28 Antigen Affinity-purified Polyclonal Antibody enhances IL-2 secretion in Jurkat human acute T cell leukemia cell line stimulated with 10 ng/mL phorbol myristate acetate (PMA) and 0.5 μM calcium ionophore, in a dose-dependent manner, as measured using the Quantikine Human IL-2 ELISA Kit (Catalog # D2050). The ED50 for this effect is typically 0.3‑0.6 μg/mL.
Immunocytochemistry
CD28 in Human PBMCs. CD28 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using 10 µg/mL Goat Anti-Human CD28 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF‑342‑PB) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counter­stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: CD28

CD28 and CTLA-4, together with their ligands, B7-1 and B7-2, constitute one of the dominant costimulatory pathways that regulate T and B cell responses. CD28 and CTLA-4 are structurally homologous molecules that are members of the immunoglobulin (Ig) gene superfamily. Both CD28 and CTLA-4 are composed of a single Ig V‑like extracellular domain, a transmembrane domain and an intracellular domain. CD28 and CTLA-4 are both expressed on the cell surface as disulfide-linked homodimers or as monomers. The genes encoding these two molecules are closely linked on human chromosome 2 and mouse chromosome 1. Mouse CD28 is expressed constitutively on virtually 100% of mouse T cells and on developing thymocytes. Cell surface expression of mouse CD28 is down-regulated upon ligation of CD28 in the presence of PMA or PHA. In contrast, CTLA-4 is not expressed constitutively but is up-regulated rapidly following T cell activation and CD28 ligation. Cell surface expression of mouse CTLA-4 peaks approximately 48 hours after activation. Although both CTLA-4 and CD28 can bind to the same ligands, CTLA-4 binds to B7-1 and B7-2 with a 20-100 fold higher affinity than CD28. CD28/B7 interaction has been shown to prevent apoptosis of activated T cells via the upregulation of Bcl-xL. CD28 ligation has also been shown to regulate Th1/Th2 differentiation.

  • References:
    1. Lenschow, D.J. et al. (1996) Annu. Rev. Immunol. 14:233.
    2. Hathcock, K.S. and R.J. Hodes (1996) Advances in Immunol. 62:131.
    3. Ward, S.G. (1996) Biochem. J. 318:361.
  • Entrez Gene IDs:
    940 (Human); 12487 (Mouse); 102146670 (Cynomolgus Monkey)
  • Alternate Names:
    CD28 antigen (Tp44); CD28 antigen; CD28 molecule; CD28; MGC138290; T-cell-specific surface glycoprotein CD28; Tp44
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

4 Citations: Showing 1 - 4
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Species
Applications
Sample Type
  1. Phenotype, function, and differentiation potential of human monocyte subsets
    Authors: LB Boyette, C Macedo, K Hadi, BD Elinoff, JT Walters, B Ramaswami, G Chalasani, JM Taboas, FG Lakkis, DM Metes
    PLoS ONE, 2017;12(4):e0176460.
    Species: Human
    Sample Type: Whole Cells
    Application: Functional Assay
  2. Dysregulated CD46 shedding interferes with Th1-contraction in systemic lupus erythematosus
    Authors: U Ellinghaus, A Cortini, CL Pinder, GL Friec, C Kemper, TJ Vyse
    Eur. J. Immunol., 2017;0(0):.
    Species: Human
    Sample Type: Whole Cells
    Application: Functional Assay
  3. The soluble forms of CD28, CD86 and CTLA-4 constitute possible immunological markers in patients with abdominal aortic aneurysm.
    Authors: Sakthivel P, Shively V, Kakoulidou M
    J. Intern. Med., 2007;261(4):399-407.
    Species: Human
    Sample Type: Plasma
    Application: ELISA Development
  4. CD28-mediated regulation of multiple myeloma cell proliferation and survival.
    Authors: Bahlis NJ, King AM, Kolonias D, Carlson LM, Liu HY, Hussein MA, Terebelo HR, Byrne GE, Levine BL, Boise LH, Lee KP
    Blood, 2007;109(11):5002-10.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC
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