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Human CD28 Antibody

Catalog #: AF-342-PB
8 Citations 2 Reviews Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF-342-PB
AF-342-SP
Human CD28 Antibody Enhances IL-2 Secretion in Jurkat Cells.
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Product Details
Citations (8)
FAQs
Supplemental Products
Reviews (2)
Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human CD28 Antibody Summary

Species Reactivity
Human
Specificity
Detects human CD28 in direct ELISAs and Western blots.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
S. frugiperda insect ovarian cell line Sf 21-derived recombinant human CD28
Asn19-Pro152
Accession # P10747
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or 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
Flow Cytometry
2.5 µg/106 cells
Human whole blood CD3+ T cells
Immunohistochemistry
3-15 µg/mL
See below
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.

Scientific Data

Agonist Activity View Larger

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 antibody in Human PBMCs by Immunocytochemistry (ICC). View Larger

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 counter-stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Immunohistochemistry CD28 antibody in Human Tonsil by Immunohistochemistry (IHC-P). View Larger

CD28 in Human Tonsil. CD28 was detected in immersion fixed paraffin-embedded sections of human tonsil using Goat Anti-Human CD28 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-342-PB) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell surfaces in lymphocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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

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Citations for Human CD28 Antibody

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.

8 Citations: Showing 1 - 8
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  1. The homodimer interfaces of costimulatory receptors B7 and CD28 control their engagement and pro-inflammatory signaling
    Authors: Popugailo, A;Rotfogel, Z;Levy, M;Turgeman, O;Hillman, D;Levy, R;Arad, G;Shpilka, T;Kaempfer, R;
    Journal of biomedical science
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  2. Mesangial Cells Exhibit Features of Antigen-Presenting Cells and Activate CD4+ T Cell Responses
    Authors: H Yu, S Cui, Y Mei, Q Li, L Wu, S Duan, G Cai, H Zhu, B Fu, L Zhang, Z Feng, X Chen
    J Immunol Res, 2019-06-17;2019(0):2121849.
    Species: Human
    Sample Types: Whole Cells
    Applications: Cell Culture
  3. Diversification of Bw4 Specificity and Recognition of a Nonclassical MHC Class I Molecule Implicated in Maternal-Fetal Tolerance by Killer Cell Ig-like Receptors of the Rhesus Macaque
    Authors: P Banerjee, M Ries, SK Janaka, AG Grandea, R Wiseman, DH O'Connor, TG Golos, DT Evans
    J. Immunol., 2018-09-19;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  4. 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., 2017-05-22;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Functional Assay
  5. 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, 2017-04-26;12(4):e0176460.
    Species: Human
    Sample Types: Whole Cells
    Applications: Functional Assay
  6. Binding of Superantigen Toxins into the CD28 Homodimer Interface Is Essential for Induction of Cytokine Genes That Mediate Lethal Shock
    Authors: Gila Arad, Revital Levy, Iris Nasie, Dalia Hillman, Ziv Rotfogel, Uri Barash et al.
    PLoS Biology
  7. 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., 2007-04-01;261(4):399-407.
    Species: Human
    Sample Types: Plasma
    Applications: ELISA Development
  8. 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, 2007-02-20;109(11):5002-10.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC

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Human CD28 Antibody
By Anonymous on 04/11/2022
Application: Immunocytochemistry/Immunofluorescence Sample Tested: Melanoma tissue Species: Human
Immunocytochemistry/Immunofluorescence Human CD28 Antibody AF-342-PB
Human CD28 Antibody
By Anonymous on 02/16/2019
Application: Func Sample Tested: Peripheral blood T cells Species: Human

96-well plates were coated with anti-CD3e -/+ AF-342 and incubated with isolated CD8+ T-cells from human PBMCs. Activation was read through intracellular interferon.

Functional Assay Human CD28 Antibody AF-342-PB