Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Western Blot, Immunocytochemistry, Agonist Activity

Cited:

Western Blot, Neutralization, Flow Cytometry, Immunocytochemistry, Binding Assay, Bioassay, Cell Culture, ELISA Development, In vitro assay, Functional Assay, Polarization, Stimulation, T Cell Activation, Tissue Culture

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 37407
View all CD28 Primary Antibodies »

Product Specifications

Immunogen

S. frugiperda insect ovarian cell line Sf 21-derived recombinant human CD28
Asn19-Pro152
Accession # P10747

Specificity

Detects human CD28 in Western blots. In Western blots, this antibody does not cross-react with recombinant mouse (rm) CD28, rhCTLA4, or rmCTLA4.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human CD28 Antibody

Human CD28 Antibody Enhances IL-2 Secretion antibody in Jurkat Cells.

Human CD28 Antibody Enhances IL-2 Secretion in Jurkat Cells.

Human CD28 Monoclonal Antibody enhances IL-2 secretion in the 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 (D2050). The ED50 for this effect is typically 0.2-0.6 µg/mL.

Detection of CD28 in U266 human myeloma cell line (Positive) & K562 human chronic myelogenous leukemia cell line (Negative).

CD28 was detected in immersion fixed U266 human myeloma cell line (Positive) & K562 human chronic myelogenous leukemia cell line (Negative) using Mouse Anti-Human CD28 Monoclonal Antibody (Catalog # MAB342) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to Cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Applications for Human CD28 Antibody

Application
Recommended Usage

Agonist Activity

0.2-0.6 µg/mL
Sample: Jurkat human acute T cell leukemia cell line

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed U266 human myeloma cell line (Positive) & K562 human chronic myelogenous leukemia cell line (Negative)

Western Blot

1 µg/mL
Sample: Recombinant Human CD28 Fc Chimera (Catalog # 342-CD)
under non-reducing conditions only

Reviewed Applications

Read 2 reviews rated 5 using MAB342 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from ascites

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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.

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.

Calculators

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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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.

Alternate Names

CD28

Entrez Gene IDs

940 (Human); 12487 (Mouse); 102146670 (Cynomolgus Monkey)

Gene Symbol

CD28

UniProt

P10747 (Human)

Additional CD28 Products

Product Documents for Human CD28 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human CD28 Antibody

For research use only

Citations for Human CD28 Antibody

Customer Reviews for Human CD28 Antibody (2)

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2 Customer Ratings
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  • Human CD28 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: T cells
    Species: Human
    Verified Customer | Posted 02/10/2022
    Human T cells
    Human CD28 Antibody MAB342
  • Human CD28 Antibody
    Name: Anonymous
    Application: Functional Assay
    Sample Tested: Peripheral blood T cells
    Species: Human
    Verified Customer | Posted 02/16/2019
    96-well plates were coated with anti-CD3e -/+ MAB342 and incubated with isolated CD8+ T-cells from human PBMCs. Activation was read through intracellular interferon.
    Human CD28 Antibody MAB342

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Protocols

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