Purified NA/LE Mouse Anti-Human CD28
Clone CD28.2 (RUO)
- Brand BD Pharmingen™
- Alternative Name CD28 antigen; T44; Tp44; TP44
- Concentration 1.0 mg/ml
- Isotype Mouse C3H x BALB/c IgG1, κ
- Reactivity Human (QC Testing)
Flow cytometry (Routinely Tested)
Bioassay (Tested During Development)
- Immunogen Human CD28 Transfected Cell Line
- Workshop No. V 5T CD28.05
- Entrez Gene ID 940
- Storage Buffer No azide/low endotoxin: Aqueous buffered solution containing no preservative, 0.2µm sterile filtered. Endotoxin level is ≤0.01 EU/µg (≤0.001 ng/µg) of protein as determined by the LAL assay.
- Regulatory Status RUO
Regulatory Status Legend
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.
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Preparation and Storage
Store undiluted at 4°C.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
This preparation contains no preservatives, thus it should be handled under aseptic conditions.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
- Ficoll-Paque is a trademark of Amersham Biosciences Limited.
- This product is sold under license. Purchase of this product does not include rights to (i) incorporate this product into the purchaser's own products for resale to end-users, or (ii) use this product to conduct for-profit research for or on behalf of another party. For information on obtaining a license to this product for such prohibited uses, contact INSERM, 7 rue Watt, 75013 Paris. Telephone: +33 1 55 03 01 60. Facsimile: +33 1 55 03 01 18. Email: email@example.com
Suggested protocol for functional studies using CD28.2 mAb:
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.