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buv395_cd86_lps_overlay.png

Flow cytometric analysis of CD86 expression on resting or activated mouse splenocytes. Freshly isolated (Left Panel) or 72-hour lipopolysaccharide-stimulated (Right Panel) mouse splenic leucocytes were pretreated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with either BD Horizon™ BUV395 Rat IgG2a, κ Isotype Control (Cat. No. 563556; dashed line histograms) or BD Horizon BUV395 Rat Anti-Mouse CD86 antibody (Cat. No. 564199; solid line histograms). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable resting (Left Panel) or activated (Right Panel) lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

564199Image1.png

Flow cytometric analysis of CD86 expression on resting or activated mouse splenocytes. Freshly isolated (Left Panel) or 72-hour lipopolysaccharide-stimulated (Right Panel) mouse splenic leucocytes were pretreated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with either BD Horizon™ BUV395 Rat IgG2a, κ Isotype Control (Cat. No. 563556; dashed line histograms) or BD Horizon BUV395 Rat Anti-Mouse CD86 antibody (Cat. No. 564199; solid line histograms). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable resting (Left Panel) or activated (Right Panel) lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

buv395_cd86_lps_overlay.png 564199Image1.png Violet-Cap_UV.png

Flow cytometric analysis of CD86 expression on resting or activated mouse splenocytes. Freshly isolated (Left Panel) or 72-hour lipopolysaccharide-stimulated (Right Panel) mouse splenic leucocytes were pretreated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with either BD Horizon™ BUV395 Rat IgG2a, κ Isotype Control (Cat. No. 563556; dashed line histograms) or BD Horizon BUV395 Rat Anti-Mouse CD86 antibody (Cat. No. 564199; solid line histograms). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable resting (Left Panel) or activated (Right Panel) lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Flow cytometric analysis of CD86 expression on resting or activated mouse splenocytes. Freshly isolated (Left Panel) or 72-hour lipopolysaccharide-stimulated (Right Panel) mouse splenic leucocytes were pretreated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with either BD Horizon™ BUV395 Rat IgG2a, κ Isotype Control (Cat. No. 563556; dashed line histograms) or BD Horizon BUV395 Rat Anti-Mouse CD86 antibody (Cat. No. 564199; solid line histograms). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable resting (Left Panel) or activated (Right Panel) lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Product Details
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BD Horizon™
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)
0.2 mg/ml
AB_2738664
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon BUV395 under optimum conditions, and unconjugated antibody and free BD Horizon BUV395 were removed.
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Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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564199 Rev. 1
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.

The antibody was conjugated to BD Horizon™ BUV395 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is optimal for multicolor flow cytometry because it has little to no spillover into other detectors. With an Ex Max at 348 nm and an Em Max at 395 nm, BD Horizon BUV395 can be excited with a 355 nm laser and detected with a 379/28 filter.

564199 Rev. 1
Format Details
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BUV395
The BD Horizon Brilliant™ Ultraviolet 395 (BUV395) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This base dye is a polymer fluorochrome with an excitation maximum (Ex Max) of 348-nm and an emission maximum (Em Max) at 395-nm. Driven by BD innovation, BUV395 is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 380-nm (e.g., 379/28-nm bandpass filter). BUV395 is the ideal dye when using only one detector on the ultraviolet laser as it spills into no other detectors and no other fluors spill into it. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BUV395
348 nm
395 nm
564199 Rev.1
Citations & References
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View product citations for antibody "564199" on CiteAb

Development References (14)

  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: Flow cytometry, Inhibition). 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: Functional assay, Immunohistochemistry, Inhibition). 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. (Clone-specific: Flow cytometry). 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. 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
  14. 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 (14) View Less
564199 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.