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Purified Hamster Anti-Mouse CD152
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).
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 (Tested During Development)
0.5 mg/ml
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.

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).

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. 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.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. An isotype control should be used at the same concentration as the antibody of interest.
553719 Rev. 12
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. 12
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.12
Citations & References
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Development References (17)

  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. (Biology). 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). View Reference
  3. Harper K, Balzano C, Rouvier E, Mattei MG, Luciani MF, Golstein P. CTLA-4 and CD28 activated lymphocyte molecules are closely related in both mouse and human as to sequence, message expression, gene structure, and chromosomal location. J Immunol. 1991; 147(3):1037-1044. (Biology). View Reference
  4. 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. (Biology). View Reference
  5. June CH, Bluestone JA, Nadler LM, Thompson CB. The B7 and CD28 receptor families. Immunol Today. 1994; 15(7):321-331. (Biology). View Reference
  6. 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. (Biology). View Reference
  7. 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
  8. Linsley PS, Bradshaw J, Greene J, Peach R, Bennett KL, Mittler RS. Intracellular trafficking of CTLA-4 and focal localization towards sites of TCR engagement. Immunity. 1996; 4(6):535-543. (Biology). View Reference
  9. Marengere LE, Waterhouse P, Duncan GS, Mittrucker HW, Feng GS, Mak TW. Regulation of T cell receptor signaling by tyrosine phosphatase SYP association with CTLA-4. Science. 1996; 272(5265):1170-1173. (Biology). View Reference
  10. 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. (Biology). View Reference
  11. Perkins D, Wang Z, Donovan C, et al. Regulation of CTLA-4 expression during T cell activation. J Immunol. 1996; 156(11):4154-4159. (Biology). View Reference
  12. 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. (Biology). View Reference
  13. 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. (Biology). View Reference
  14. Tivol EA, Boyd SD, McKeon S, et al. CTLA4Ig prevents lymphoproliferation and fatal multiorgan tissue destruction in CTLA-4-deficient mice. J Immunol. 1997; 158(11):5091-5094. (Biology). View Reference
  15. 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
  16. Waterhouse P, Bachmann MF, Penninger JM, Ohashi PS, Mak TW. Normal thymic selection, normal viability and decreased lymphoproliferation in T cell receptor-transgenic CTLA-4-deficient mice. Eur J Immunol. 1997; 27(8):1887-1892. (Biology). View Reference
  17. Waterhouse P, Penninger JM, Timms E, et al. Lymphoproliferative disorders with early lethality in mice deficient in Ctla-4. Science. 1995; 270(5238):985-988. (Biology). View Reference
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553719 Rev. 12

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