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Biotin Hamster Anti-Mouse CD28
Biotin Hamster Anti-Mouse CD28

Two-color analysis of CD28 expression on splenic T lymphocytes. After pre-incubation with purified anti-mouse CD16/CD32 mAb 2.4G2 [Mouse BD Fc Block™] (Cat. No. 553141), BALB/c splenocytes were simultaneously stained with FITC-conjugated anti-mouse CD4 mAb RM4-5 (Cat. No. 553046) and FITC-conjugated anti-mouse CD8a mAb 53-6.7 (Cat. No. 553030), in addition to staining with biotinylated anti-mouse CD28 clone 37.51 (right panel), followed by streptavidin-PE (Cat. No. 554061). Flow cytometry was performed on a BD FACScan™ flow cytometry system.

Two-color analysis of CD28 expression on splenic T lymphocytes. After pre-incubation with purified anti-mouse CD16/CD32 mAb 2.4G2 [Mouse BD Fc Block™] (Cat. No. 553141), BALB/c splenocytes were simultaneously stained with FITC-conjugated anti-mouse CD4 mAb RM4-5 (Cat. No. 553046) and FITC-conjugated anti-mouse CD8a mAb 53-6.7 (Cat. No. 553030), in addition to staining with biotinylated anti-mouse CD28 clone 37.51 (right panel), followed by streptavidin-PE (Cat. No. 554061). Flow cytometry was performed on a BD FACScan™ flow cytometry system.

Product Details
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BD Pharmingen™
Mouse (QC Testing)
Syrian Hamster IgG2, λ1
Mouse EL-4 (T-cell lymphoma) Cells
Flow cytometry (Routinely Tested)
0.5 mg/ml
12487
AB_394765
Aqueous buffered solution containing protein stabilizer and ≤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 with biotin under optimum conditions, and unreacted biotin was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

Flow Cytometry: For flow cytometry of cell suspensions from peripheral lymphoid tissues, it is recommended that multicolor staining be performed to identify T lymphocytes and/or NK cells and that purified anti-mouse CD16/CD32 mAb 2.4G2 [Mouse BD Fc Block™] (Cat. No. 553141) be used. Since this antigen is expressed at low density on resting peripheral T lymphocytes, it is recommended that a "bright" second-step reagent, such as Streptavidin-PE (Cat. No. 554061), be used for flow cytometric staining of these cell populations.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. A table of the hamster IgG groups, Reactivity of Mouse Anti-Hamster Ig mAbs, may be viewed at http://www.bdbiosciences.com/documents/hamster_chart_11x17.pdf.
  4. 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.
553296 Rev. 15
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.

This antibody is routinely tested by flow cytometric analysis. Other applications were tested at BD Biosciences Pharmingen during antibody development only or reported in the literature.

553296 Rev. 15
Format Details
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Biotin
Biotin is a ubiquitous co-factor (also known as Vitamin B7) that has many properties that make it extremely useful for molecular biology. Biotin has an extremely high affinity for the Avidin family of proteins (Kd = 10-15 M), making it the perfect tool to link two molecules. Biotin labeled antibodies can be combined with any number of Avidin-conjugated probes in order to customize an assay to a particular need. This is especially useful in the case of magnetic cell separation using streptavidin/magnetic bead conjugates, or in the case of flow cytometry using streptavidin/fluorophore conjugates.
Biotin
553296 Rev.15
Citations & References
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Development References (15)

  1. 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. (Biology). View Reference
  2. 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. (Biology). View Reference
  3. 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, Immunoprecipitation). View Reference
  4. 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. (Biology). View Reference
  5. 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. (Biology). View Reference
  6. June CH, Bluestone JA, Nadler LM, Thompson CB. The B7 and CD28 receptor families. Immunol Today. 1994; 15(7):321-331. (Biology). View Reference
  7. 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. (Biology). View Reference
  8. 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. (Biology). View Reference
  9. 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. (Biology). View Reference
  10. 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. (Biology). View Reference
  11. 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
  12. 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. (Biology). View Reference
  13. 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. (Biology). View Reference
  14. 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
  15. 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. (Biology). View Reference
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553296 Rev. 15

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