CD28 antibody

Catalog No. ABIN1177331

The Mouse Monoclonal anti-CD28 antibody has been validated for FACS and BCA. It is suitable to detect CD28 in samples from Human.

Quick Overview for CD28 antibody (ABIN1177331)

Target: CD28

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This CD28 antibody is un-conjugated

Application: Flow Cytometry (FACS), Biochemical Assay (BCA)

Clone: CD28-2

Product Details anti-CD28 Antibody

Brand: BD Pharmingen™

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Sterility: 0.2 μm filtered

Endotoxin Level: Endotoxin level is ≤ 0.01 EU/μg (≤ 0.001 ng/μg) of protein as determined by the LAL assay.

Immunogen: Human CD28 Transfected Cell Line

Isotype: IgG1 kappa

Application Details

Protocol: 1. Isolate human PBMC with Ficoll-Paque™ Plus
2. Suspend PBMC in medium supplemented with 1% glutamine, 1% penicillin/streptomycin and 10% FBS at 10^6 cells/ml
3. Incubate 1 x 10^6 PBMC with soluble NA/LE™ format of CD3 mAb (Cat. No. 555336) at 1µg/ml final concentration in 7% CO2, 37°C for 3 days
4. Wash cells twice and resuspend in the above medium at 10^6 cells/ml
5. Distribute cells in 96-well round bottom plate at 10^5 cells/well
6. Add soluble NA/LE™ format of CD28.2 mAb at 5 µg/ml final concentration
7. Incubate the plate for an additional three days 8. Pulse the plate with [3H]TdR (1 µCi/well). Harvest the plate 20 hours after pulse and count
Comment: Addition of Protein G (Sigma) 5 µg/ml together with CD28.2 will significantly enhance the costimulation, presumably through cross-linking.

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 1.0 mg/mL

Buffer: No azide/low endotoxin: Aqueous buffered solution containing no preservative, 0.2μm sterile filtered.

Preservative: Azide free

Storage: 4 °C

Storage Comment: Store undiluted at 4°C. This preparation contains no preservatives, thus it should be handled under aseptic conditions.

Target Details for CD28

Target: CD28

Alternative Name: CD28

Background: The CD28.2 monoclonal antibody specifically binds to CD28, a 44 kDa homodimeric transmembrane glycoprotein present on most mature T cells, thymocytes and plasma cells. CD28 is a costimulatory receptor that binds CD80 and CD86 as ligands and plays a very important role in T cell-B cell interactions. It has been suggested that CD28 initiates and regulates a separate and distinct signal transduction pathway from those stimulated by the TCR complex. Additionally, it has been reported that CD28 antibody clones vary in their ability to stimulate T cells to produce IL-2 and increase intracellular Ca2+ concentration. This finding suggests the existence of functionally distinct subregions on the CD28 molecule. CD28.2 has been demonstrated to bind to the same molecule as clone L293, another CD28 mAb, and has been reported to induce Ca2+ influx in Jurkat T cells.
Synonyms: CD28 antigen, T44, Tp44, TP44

Pathways: TCR Signaling, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response