Preparation And Storage
Recommended Assay Procedures
For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. Diagnostic uses require a separate license.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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: techtransfert@inserm-transfert.fr
Companion Products
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.
The antibody was conjugated to BD Horizon BB515 which was developed exclusively by BD Biosciences. With an excitation max of 490 nm and an emission max of 515 nm, BD Horizon BB515 can be excited by the 488 nm laser and detected in a standard FITC set (eg, 530/30-nm filter). This dye provides a much brighter alternative to FITC with less spillover into the PE detector.
Development References (6)
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Barclay NA, Brown MH, Birkeland ML, et al, ed. The Leukocyte Antigen FactsBook. San Diego, CA: Academic Press; 1997.
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June CH, Bluestone JA, Nadler LM, Thompson CB. The B7 and CD28 receptor families. Immunol Today. 1994; 15(7):321-331. (Biology). View Reference
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Kuiper H, Brouwer M, Vermeire S, van Lier R. Analysis of the Workshop CD28 Panel mAb: distinct signalling pathways coupled to CD28. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:373-374.
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Nunes J, Klasen S, Franco MD, et al. Signalling through CD28 T-cell activation pathway involves an inositol phospholipid-specific phospholipase C activity. Biochem J. 1993; 293(3):835-842. (Clone-specific: Activation, Calcium Flux, (Co)-stimulation, Functional assay). View Reference
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Nunes J, Klasen S, Ragueneau M, et al. CD28 mAbs with distinct binding properties differ in their ability to induce T cell activation: analysis of early and late activation events. Int Immunol. 1993; 5(3):311-315. (Immunogen: Activation, Calcium Flux, (Co)-stimulation, Flow cytometry, Functional assay, IC/FCM Block, Immunoprecipitation, Stimulation). View Reference
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Olive D, Cerdan C, Costello R, et al. CD28 and CTLA-4 cluster report. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:360-370.
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.