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09274D_553691_image2.png

Upregulation of membrane CD86 (B7-2) on activated B lymphocytes. Freshly isolated (left panel) or 72-hour LPS-stimulated BALBc splenocytes (right panel) were pretreated with Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141/553142) and either stained with FITC-conjugated GL1 mAb (open histograms) or unstained (shaded histograms). Flow cytometry was performed on a BD FACScan™ Flow Cytometry System. Resting lymphocytes (left panel) or activated blasts (right panel) were selected according to light-scatter profile.

553691Image1.png

Upregulation of membrane CD86 (B7-2) on activated B lymphocytes. Freshly isolated (left panel) or 72-hour LPS-stimulated BALBc splenocytes (right panel) were pretreated with Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141/553142) and either stained with FITC-conjugated GL1 mAb (open histograms) or unstained (shaded histograms). Flow cytometry was performed on a BD FACScan™ Flow Cytometry System. Resting lymphocytes (left panel) or activated blasts (right panel) were selected according to light-scatter profile.

09274D_553691_image2.png 553691Image1.png Green-Cap-Blue-Label.png

Upregulation of membrane CD86 (B7-2) on activated B lymphocytes. Freshly isolated (left panel) or 72-hour LPS-stimulated BALBc splenocytes (right panel) were pretreated with Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141/553142) and either stained with FITC-conjugated GL1 mAb (open histograms) or unstained (shaded histograms). Flow cytometry was performed on a BD FACScan™ Flow Cytometry System. Resting lymphocytes (left panel) or activated blasts (right panel) were selected according to light-scatter profile.

Upregulation of membrane CD86 (B7-2) on activated B lymphocytes. Freshly isolated (left panel) or 72-hour LPS-stimulated BALBc splenocytes (right panel) were pretreated with Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141/553142) and either stained with FITC-conjugated GL1 mAb (open histograms) or unstained (shaded histograms). Flow cytometry was performed on a BD FACScan™ Flow Cytometry System. Resting lymphocytes (left panel) or activated blasts (right panel) were selected according to light-scatter profile.

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)
0.5 mg/ml
AB_394993
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 with FITC under optimum conditions, and unreacted FITC was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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Recommended Assay Procedures

Mouse BD Fc Block™ purified anti-mouse CD16/32 mAb 2.4G2 (Cat. No. 553141/553142) may help to reduce non-specific binding of GL1 antibody to cells bearing Fcγ-receptors.

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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.
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553691 Rev. 14
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.

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.

553691 Rev. 14
Format Details
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FITC
Fluorescein (FITC) is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 518-nm. FITC is designed to be excited by the Blue laser (488-nm) and detected using an optical filter centered near 520 nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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FITC
494 nm
518 nm
553691 Rev.14
Citations & References
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View product citations for antibody "553691" on CiteAb

Development References (17)

  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, Immunofluorescence). View Reference
View All (17) View Less
553691 Rev. 14

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.