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Biotin Rat Anti-Mouse CD86
Biotin Rat Anti-Mouse CD86

Fow cytometric analysis of  CD86 expression of activated and resting mouse splenocytes. Freshly isolated (Left Panel) or 72-hour LPS-stimulated BALB/c splenic luecocytes (Right Panel) were pretreated with Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) (Cat. No. 553141/553142).   The cells were then either unstained (shaded histograms) or stained with Biotin Rat Anti-Mouse CD86 (Cat. No 553690); open histograms) followed by Avidin-FITC (Cat. No. 554057; shaded and open histograms). Flow cytometry was performed on a BD FACScan™ Flow Cytometry System. The fluorescence histograms were derived from gated events with the forward and side light-catter characteristics of viable resting (Left Panel) or activated (Right Panel) lymphocytes.

Fow cytometric analysis of  CD86 expression of activated and resting mouse splenocytes. Freshly isolated (Left Panel) or 72-hour LPS-stimulated BALB/c splenic luecocytes (Right Panel) were pretreated with Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) (Cat. No. 553141/553142).   The cells were then either unstained (shaded histograms) or stained with Biotin Rat Anti-Mouse CD86 (Cat. No 553690); open histograms) followed by Avidin-FITC (Cat. No. 554057; shaded and open histograms). Flow cytometry was performed on a BD FACScan™ Flow Cytometry System. The fluorescence histograms were derived from gated events with the forward and side light-catter characteristics of viable resting (Left Panel) or activated (Right Panel) lymphocytes.

Product Details
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BD Pharmingen™
B7-2; Ly-58; Cd28l2; Early T-cell costimulatory molecule 1; ETC1; MB7; CLS1
Mouse (QC Testing)
Rat LOU, also known as Louvain, LOU/C, LOU/M IgG2a, κ
Mouse (CBA/Ca) LPS-activated splenic B Cells
Flow cytometry (Routinely Tested), Immunohistochemistry-frozen (Reported)
0.5 mg/ml
12524
AB_394992
Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. 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.

Recommended Assay Procedures

Immunofluorescent staining: The use of Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) (Cat. No. 553141/553142) may help to reduce non-specific binding of GL1 antibody to cells bearing Fcγ-receptors.

Immunohistochemistry: For IHC, we recommend the use of Purified Rat Anti-Mouse CD86 (Clone GL1, Cat. No. 550542) which has been formulated specifically for immunohistochemistry.

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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
553690 Rev. 17
Antibody Details
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GL1

The GL1 antibody specifically recognizes the B7-2 (CD86) costimulatory molecule expressed on a broad spectrum of leukocytes, including B lymphocytes, T lymphocytes, thioglycollate-induced peritoneal macrophages, dendritic cells and astrocytes. CD86 is expressed at low levels by freshly explanted peripheral B and T cells, and its expression is substantially increased by a variety of T cell- and B cell-specific stimuli with a peak expression after 18-42 hours of culture. In contrast to most naive CD4+ T cells, memory CD4+ T cells express B7-2, both at the mRNA and protein level. CD86, a ligand for CD28 and CD152 (CTLA-4), is one of the accessory molecules that plays an important role in T cell-B cell costimulatory interactions. It has been shown to be involved in immunoglobulin class-switching and triggering of mouse NK cell-mediated cytotoxicity. CD80 (B7-1) is an alternate ligand for CD28 and CD152 (CTLA-4). GL1 antibody reportedly blocks MLR and stimulation of T cells by natural antigen-presenting cells. In addition, a mixture of anti-B7-1 and anti B7-2 (GL1) mAbs reportedly inhibits the in vitro interaction of CTLA-4 with its ligand and the in vivo priming of cytotoxic T lymphocytes.

553690 Rev. 17
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
553690 Rev.17
Citations & References
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Development References (18)

  1. Bluestone JA. New perspectives of CD28-B7-mediated T cell costimulation. Immunity. 1995; 2(6):555-559. (Biology). View Reference
  2. Borriello F, Sethna MP, Boyd SD, et al. B7-1 and B7-2 have overlapping, critical roles in immunoglobulin class switching and germinal center formation. Immunity. 1997; 6(3):303-313. (Biology). View Reference
  3. Freeman GJ, Borriello F, Hodes RJ, et al. Uncovering of functional alternative CTLA-4 counter-receptor in B7-deficient mice. Science. 1993; 262(5135):907-909. (Biology). View Reference
  4. Hakamada-Taguchi R, Kato T, Ushijima H, Murakami M, Uede T, Nariuchi H. Expression and co-stimulatory function of B7-2 on murine CD4+ T cells. Eur J Immunol. 1998; 28(3):865-873. (Biology). View Reference
  5. Hathcock KS, Laszlo G, Dickler HB, Bradshaw J, Linsley P, Hodes RJ. Identification of an alternative CTLA-4 ligand costimulatory for T cell activation. Science. 1993; 262(5135):905-907. (Immunogen: Blocking, Immunoprecipitation). View Reference
  6. Hathcock KS, Laszlo G, Pucillo C, Linsley P, Hodes RJ. Comparative analysis of B7-1 and B7-2 costimulatory ligands: expression and function. J Exp Med. 1994; 180(2):631-640. (Clone-specific: Blocking). View Reference
  7. Inaba K, Witmer-Pack M, Inaba M, et al. The tissue distribution of the B7-2 costimulator in mice: abundant expression on dendritic cells in situ and during maturation in vitro. J Exp Med. 1994; 180(5):1849-1860. (Clone-specific: Blocking, Immunohistochemistry). 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: Blocking). View Reference
  9. Larsen CP, Ritchie SC, Hendrix R, et al. Regulation of immunostimulatory function and costimulatory molecule (B7-1 and B7-2) expression on murine dendritic cells. J Immunol. 1994; 152(11):5208-5219. (Biology). View Reference
  10. Lenschow DJ, Su GH, Zuckerman LA, et al. Expression and functional significance of an additional ligand for CTLA-4. Proc Natl Acad Sci U S A. 1993; 90(23):11054-11058. (Biology). View Reference
  11. Liu Y, Wenger RH, Zhao M, Nielsen PJ. Distinct costimulatory molecules are required for the induction of effector and memory cytotoxic T lymphocytes. J Exp Med. 1997; 185(2):251-262. (Clone-specific: Blocking). View Reference
  12. Martin-Fontecha A, Assarsson E, Carbone E, Karre K, Ljunggren HG. Triggering of murine NK cells by CD40 and CD86 (B7-2). J Immunol. 1999; 162(10):5910-5916. (Biology). View Reference
  13. McAdam AJ, Schweitzer AN, Sharpe AH. The role of B7 co-stimulation in activation and differentiation of CD4+ and CD8+ T cells. Immunol Rev. 1998; 165:231-247. (Biology). View Reference
  14. Nikcevich KM, Gordon KB, Tan L, et al. IFN-gamma-activated primary murine astrocytes express B7 costimulatory molecules and prime naive antigen-specific T cells. J Immunol. 1997; 158(2):614-621. (Biology). View Reference
  15. Rauschmayr-Kopp T, Williams IR, Borriello F, Sharpe AH, Kupper TS. Distinct roles for B7 costimulation in contact hypersensitivity and humoral immune responses to epicutaneous antigen. Eur J Immunol. 1998; 28(12):4221-4227. (Biology). View Reference
  16. Roy M, Aruffo A, Ledbetter J, Linsley P, Kehry M, Noelle R. Studies on the interdependence of gp39 and B7 expression and function during antigen-specific immune responses. Eur J Immunol. 1995; 25(2):596-603. (Biology). View Reference
  17. Turley SJ, Inaba K, Garrett WS, et al. Transport of peptide-MHC class II complexes in developing dendritic cells. Science. 2000; 288(5465):522-527. (Clone-specific: Electron microscopy, Fluorescence microscopy). View Reference
  18. Yang G, Mizuno MT, Hellstrom KE, Chen L. B7-negative versus B7-positive P815 tumor: differential requirements for priming of an antitumor immune response in lymph nodes. J Immunol. 1997; 158(2):851-858. (Clone-specific: Immunohistochemistry). View Reference
View All (18) View Less
553690 Rev. 17

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