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      RB705 Hamster Anti-Mouse CD28
      RB705 Hamster Anti-Mouse CD28
      Multiparameter flow cytometric analysis using BD OptiBuild™ RB705 Hamster Anti-Mouse CD28 antibody (Cat. No. 756832; Right Plot) on viable C57BL/6 Mouse splenocytes, with Autofluorescence Control (Left Plot). Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer.
      RB705 Hamster Anti-Mouse CD28
      Multiparameter flow cytometric analysis using BD OptiBuild™ RB705 Hamster Anti-Mouse CD28 antibody (Cat. No. 756832; Right Plot) on viable C57BL/6 Mouse splenocytes, with Autofluorescence Control (Left Plot). Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer.
      RB705 Hamster Anti-Mouse CD28 RB705 Hamster Anti-Mouse CD28
      Multiparameter flow cytometric analysis using BD OptiBuild™ RB705 Hamster Anti-Mouse CD28 antibody (Cat. No. 756832; Right Plot) on viable C57BL/6 Mouse splenocytes, with Autofluorescence Control (Left Plot). Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer.
      Multiparameter flow cytometric analysis using BD OptiBuild™ RB705 Hamster Anti-Mouse CD28 antibody (Cat. No. 756832; Right Plot) on viable C57BL/6 Mouse splenocytes, with Autofluorescence Control (Left Plot). Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer.
      Product Details
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      BD OptiBuild™
      Cd28; CD28 antigen; T-cell-specific surface glycoprotein CD28
      Mouse (Tested in Development)
      Syrian Hamster IgG2, λ1
      Mouse EL-4 (T-cell lymphoma) Cells
      Flow cytometry (Qualified)
      0.2 mg/ml
      12487
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions that minimize unconjugated dye and antibody. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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      Recommended Assay Procedures

      BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

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      Product Notices

      1. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
      2. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
      3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      4. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      5. An isotype control should be used at the same concentration as the antibody of interest.
      6. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
      7. 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.
      8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      9. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
      10. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      11. For U.S. patents that may apply, see bd.com/patents.
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      756832 Rev. 0
      Antibody Details
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      37.51

      The 37.51 antibody reacts with CD28, which is expressed on most thymocytes, at low density on nearly all CD4+ and CD8+ peripheral T cells, and at even lower density on NK cells. The expression of CD28, in splenocytes and thymocytes,  has been reported to increase after activation. CD28 transcripts are found in mast cells, and cell-surface expression of CD28 is induced upon maturation or activation of mast cells. It has been reported that CD28 is not expressed on some populations of intraepithelial T lymphocytes. CD28 is a costimulatory receptor; its ligands include CD80 (B7-1) and CD86 (B7-2). The 37.51 mAb augments proliferation and cytokine production by activated T and NK cells and can provide a costimulatory signal for CTL induction. There is considerable evidence that CD28 is a costimulatory receptor involved in many, but not all, T cell-dependent immune responses.

      756832 Rev. 0
      Format Details
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      RB705
      The BD Horizon RealBlue™ 705 (RB705) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 707-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB705 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB705 can be used as an alternative to PerCP-Cy5.5 or BB700 and we recommend using an optical filter centered near 710-nm (e.g., a 695/40 or 710/50-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PerCP-Cy5.5. RB705 is on average brighter than PerCP-Cy5.5 and BB700, and has minimal spillover into Yellow-Green detectors.
      altImg
      RB705
      Blue 488 nm
      498 nm
      707 nm
      756832 Rev.0
      Citations & References
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      View product citations for antibody "756832" on CiteAb

      Development References (16)

      1. Bluestone JA. New perspectives of CD28-B7-mediated T cell costimulation. Immunity. 1995; 2(6):555-559. (Biology: Apoptosis). View Reference
      2. Cibotti R, Punt JA, Dash KS, Sharrow SO, Singer A. Surface molecules that drive T cell development in vitro in the absence of thymic epithelium and in the absence of lineage-specific signals. Immunity. 1997; 6(3):245-255. (Clone-specific: (Co)-stimulation, Stimulation). View Reference
      3. Gelfanov V, Lai YG, Gelfanova V, Dong JY, Su JP, Liao NS. Differential requirement of CD28 costimulation for activation of murine CD8+ intestinal intraepithelial lymphocyte subsets and lymph node cells. J Immunol. 1995; 155(1):76-82. (Clone-specific: (Co)-stimulation, Flow cytometry, Stimulation). View Reference
      4. Gross JA, Callas E, Allison JP. Identification and distribution of the costimulatory receptor CD28 in the mouse. J Immunol. 1992; 149(2):380-388. (Immunogen: (Co)-stimulation, Flow cytometry, Immunoprecipitation, Stimulation). View Reference
      5. Harding FA, Allison JP. CD28-B7 interactions allow the induction of CD8+ cytotoxic T lymphocytes in the absence of exogenous help. J Exp Med. 1993; 177(6):1791-1796. (Clone-specific: (Co)-stimulation, Inhibition, Stimulation). View Reference
      6. Harding FA, McArthur JG, Gross JA, Raulet DH, Allison JP. CD28-mediated signalling co-stimulates murine T cells and prevents induction of anergy in T-cell clones. Nature. 1992; 356(6370):607-609. (Clone-specific: (Co)-stimulation, Stimulation). View Reference
      7. June CH, Bluestone JA, Nadler LM, Thompson CB. The B7 and CD28 receptor families. Immunol Today. 1994; 15(7):321-331. (Biology). View Reference
      8. Krummel MF, Allison JP. CD28 and CTLA-4 have opposing effects on the response of T cells to stimulation. J Exp Med. 1995; 182(2):459-465. (Clone-specific: (Co)-stimulation, Stimulation). View Reference
      9. Lepesant H, Pierres M, Naquet P. Deficient antigen presentation by thymic epithelial cells reveals differential induction of T cell clone effector functions by CD28-mediated costimulation. Cell Immunol. 1995; 161(2):279-287. (Clone-specific: (Co)-stimulation, Stimulation). View Reference
      10. Marietta EV, Weis JJ, Weis JH. CD28 expression by mouse mast cells is modulated by lipopolysaccharide and outer surface protein A lipoprotein from Borrelia burgdorferi. J Immunol. 1997; 159(6):2840-2848. (Clone-specific: (Co)-stimulation, Flow cytometry, Stimulation). View Reference
      11. Nandi D, Gross JA, Allison JP. CD28-mediated costimulation is necessary for optimal proliferation of murine NK cells. J Immunol. 1994; 152(7):3361-3369. (Clone-specific: (Co)-stimulation, Flow cytometry, Stimulation). View Reference
      12. Nishio M, Spielman J, Lee RK, Nelson DL, Podack ER. CD80 (B7.1) and CD54 (intracellular adhesion molecule-1) induce target cell susceptibility to promiscuous cytotoxic T cell lysis. J Immunol. 1996; 157(10):4347-4353. (Biology). View Reference
      13. Ong CJ, Lim AS, Teh HS. CD28-induced cytokine production and proliferation by thymocytes are differentially regulated by the p59fyn tyrosine kinase. J Immunol. 1997; 159(5):2169-2176. (Clone-specific: (Co)-stimulation, Stimulation). View Reference
      14. Rakasz E, Hagen M, Sandor M, Lynch RG. Gamma delta T cells of the murine vagina: T cell response in vivo in the absence of the expression of CD2 and CD28 molecules. Int Immunol. 1997; 9(1):161-167. (Clone-specific: Flow cytometry). View Reference
      15. Shahinian A, Pfeffer K, Lee KP, et al. Differential T cell costimulatory requirements in CD28-deficient mice. Science. 1993; 261(5121):609-612. (Biology). View Reference
      16. Wells AD, Gudmundsdottir H, Turka LA. Following the fate of individual T cells throughout activation and clonal expansion. Signals from T cell receptor and CD28 differentially regulate the induction and duration of a proliferative response. J Clin Invest. 1997; 100(12):3173-3183. (Clone-specific: (Co)-stimulation, Stimulation). View Reference
      View All (16) View Less
      756832 Rev. 0

      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.