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      Purified Hamster Anti-Mouse CD152

      BD Pharmingen™ Purified Hamster Anti-Mouse CD152

      Clone UC10-4F10-11 (also known as UC10-4F10; 4F10)

      (RUO)
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      Purified Hamster Anti-Mouse CD152
      Expression of CD152 on activated T lymphocytes. C57BL/6 splenocytes were stained with either purified mAb UC10-4F10-11 (open histogram) or unstained (filled histogram), in the presence of Mouse Fc Block™ (purified anti-mouse CD16/CD32 mAb 2.4G2, Cat. No. 553141/553142), followed by biotinylated anti-hamster IgG cocktail (Cat. No. 554010) then Streptavidin-PE (Cat. No. 554061). The total viable leukocytes are displayed in the left panel. 48-hour ConA-activated C57BL/6 splenocytes were stained with FITC-conjugated anti-mouse CD4 mAb RM4-5 (Cat. No. 553046/553047), FITC-conjugated anti-mouse CD8a mAb 53-6.7 (Cat. No. 553030/553031), and purified mAb UC10-4F10-11 (open histogram), or no purified antibody (filled histogram), in the presence of Mouse Fc Block™, followed by biotinylated anti-hamster IgG cocktail then Streptavidin-PE. The total viable T lymphocytes (CD4+ and/or CD8a+) are displayed on the right panel. Flow cytometry was performed on a BD FACScan™ System (BD Biosciences, San Jose, CA).
      Purified Hamster Anti-Mouse CD152
      Expression of CD152 on activated T lymphocytes. C57BL/6 splenocytes were stained with either purified mAb UC10-4F10-11 (open histogram) or unstained (filled histogram), in the presence of Mouse Fc Block™ (purified anti-mouse CD16/CD32 mAb 2.4G2, Cat. No. 553141/553142), followed by biotinylated anti-hamster IgG cocktail (Cat. No. 554010) then Streptavidin-PE (Cat. No. 554061). The total viable leukocytes are displayed in the left panel. 48-hour ConA-activated C57BL/6 splenocytes were stained with FITC-conjugated anti-mouse CD4 mAb RM4-5 (Cat. No. 553046/553047), FITC-conjugated anti-mouse CD8a mAb 53-6.7 (Cat. No. 553030/553031), and purified mAb UC10-4F10-11 (open histogram), or no purified antibody (filled histogram), in the presence of Mouse Fc Block™, followed by biotinylated anti-hamster IgG cocktail then Streptavidin-PE. The total viable T lymphocytes (CD4+ and/or CD8a+) are displayed on the right panel. Flow cytometry was performed on a BD FACScan™ System (BD Biosciences, San Jose, CA).
      Product Details
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      BD Pharmingen™
      Ctla4; CTLA-4; Cytotoxic T-lymphocyte-associated protein 4; Ly-56
      Mouse (QC Testing)
      Armenian Hamster IgG1, κ
      Mouse CTLA-4 IgG2a Fusion
      Flow cytometry (Routinely Tested), Immunoprecipitation, Intracellular staining (flow cytometry) (Tested During Development)
      0.5 mg/ml
      12477
      AB_395004
      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. Store undiluted at 4°C.
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      Recommended Assay Procedures

      Since CD152 is expressed at low density on activated T cells, it may be necessary to amplify the signal by using a biotinylated second-step reagent, followed by a "bright" third-step reagent. We have found that biotin-conjugated mouse anti-hamster IgG (Cat. No. 554010) plus Streptavidin-PE (Cat. No. 554061) are effective. Mouse BD Fc Block™ (anti-mouse CD16/CD32 mAb 2.4G2, Cat. No. 553141/553142) may help to reduce non-specific binding of the antibody to cells bearing Fcγ receptors. Since a large proportion of the CTLA-4 molecule is intracellular, detection of the antigen is enhanced by staining cells permeabilized with the BD Cytofix/Cytoperm™ intracellular staining kit (Cat. No. 554714).

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      3. An isotype control should be used at the same concentration as the antibody of interest.
      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.
      5. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
      6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      7. For U.S. patents that may apply, see bd.com/patents.
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      553719 Rev. 13
      Antibody Details
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      UC10-4F10-11

      The UC10-4F10-11 monoclonal antibody specifically binds to CD152, which is also known as Cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4). CD152 is expressed on activated T lymphocytes 2-3 days after stimulation through T cell receptor. CTLA-4 has significant similarity to CD28 in amino acid sequence, structure, and genomic organization. Furthermore, CD152 and CD28 share common B7 family counter-receptors. Unlike CD28, CD152 expression appears to be restricted to activated T cells and CD25+CD4+ regulatory T (Treg) cells. Whereas CD28 delivers a costimulatory signal required for T-cell activation, CTLA-4 is a negative regulator of cell-mediated immune responses. CD152 may play roles in induction and/or maintenance of immunological tolerance, regulation of protective immunity, and autoimmune responses, and regulation of some aspects of thymocyte maturation. This hamster mAb to a mouse leukocyte antigen does not cross-react with rat leucocytes.

      553719 Rev. 13
      Format Details
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      Purified
      Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
      Purified
      553719 Rev.13
      Citations & References
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      Development References (11)

      1. Alegre ML, Noel PJ, Eisfelder BJ, et al. Regulation of surface and intracellular expression of CTLA4 on mouse T cells. J Immunol. 1996; 157(11):4762-4770. (Clone-specific: Flow cytometry, Fluorescence microscopy, Immunofluorescence, Intracellular Staining/Flow Cytometry). View Reference
      2. Cilio CM, Daws MR, Malashicheva A, Sentman CL, Holmberg D. Cytotoxic T lymphocyte antigen 4 is induced in the thymus upon in vivo activation and its blockade prevents anti-CD3-mediated depletion of thymocytes. J Exp Med. 1998; 188(7):1239-1246. (Clone-specific: Blocking, Intracellular Staining/Flow Cytometry). View Reference
      3. Issazadeh S, Zhang M, Sayegh MH, Khoury SJ.. Acquired thymic tolerance: role of CTLA4 in the initiation and maintenance of tolerance in a clinically relevant autoimmune disease model. J Immunol. 1999; 162(2):761-765. (Clone-specific: In vivo exacerbation). View Reference
      4. Kearney ER, Walunas TL, Karr RW, et al. Antigen-dependent clonal expansion of a trace population of antigen-specific CD4+ T cells in vivo is dependent on CD28 costimulation and inhibited by CTLA-4. J Immunol. 1995; 155(3):1032-1036. (Clone-specific: In vivo exacerbation). View Reference
      5. Lee KM, Chuang E, Griffin M, et al. Molecular basis of T cell inactivation by CTLA-4. Science. 1998; 282(5397):2263-2266. (Clone-specific: Immunoprecipitation). View Reference
      6. McCoy K, Camberis M, Gros GL. Protective immunity to nematode infection is induced by CTLA-4 blockade. J Exp Med. 1997; 186(2):183-187. (Clone-specific: In vivo exacerbation). View Reference
      7. Perkins D, Wang Z, Donovan C, et al. Regulation of CTLA-4 expression during T cell activation. J Immunol. 1996; 156(11):4154-4159. (Clone-specific: Flow cytometry). View Reference
      8. Perrin PJ, Maldonado JH, Davis TA, June CH, Racke MK. CTLA-4 blockade enhances clinical disease and cytokine production during experimental allergic encephalomyelitis. J Immunol. 1996; 157(4):1333-1336. (Clone-specific: In vivo exacerbation). View Reference
      9. Read S, Malmstrom V, Powrie F. Cytotoxic T lymphocyte-associated antigen 4 plays an essential role in the function of CD25(+)CD4(+) regulatory cells that control intestinal inflammation. J Exp Med. 2000; 192(2):295-302. (Clone-specific: Intracellular Staining/Flow Cytometry). View Reference
      10. Takahashi T, Tagami T, Yamazaki S, et al. Immunologic self-tolerance maintained by CD25(+)CD4(+) regulatory T cells constitutively expressing cytotoxic T lymphocyte-associated antigen 4. J Exp Med. 2000; 192(2):303-309. (Clone-specific: Flow cytometry, In vivo exacerbation). View Reference
      11. Walunas TL, Lenschow DJ, Bakker CY, et al. CTLA-4 can function as a negative regulator of T cell activation.. Immunity. 1994; 1(5):405-13. (Immunogen: Flow cytometry, Immunoprecipitation, Stimulation). View Reference
      View All (11) View Less
      553719 Rev. 13

      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.

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