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BB700 Rat Anti-Mouse CD86
Product Details
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BD OptiBuild™
B7-2; Ly-58; Cd28l2; Early T-cell costimulatory molecule 1; ETC1; MB7; CLS1
Mouse (Tested in Development)
Rat LOU, also known as Louvain, LOU/C, LOU/M IgG2a, κ
Mouse (CBA/Ca) LPS-activated splenic B Cells
Flow cytometry (Qualified)
0.2 mg/ml
12524
AB_2871388
Aqueous buffered solution containing ≤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 BB700 under optimal conditions that minimize unconjugated dye and antibody.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794 or 566349).

When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values.

For optimal results, it is recommended to perform two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent staining prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

Product Notices

  1. This antibody was developed for use in flow cytometry.
  2. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
  3. Researchers should determine the optimal concentration of this reagent for their individual applications.
  4. An isotype control should be used at the same concentration as the antibody of interest.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  8. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  9. BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
  10. Cy is a trademark of GE Healthcare.
742120 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™ BB700, which is part of the BD Horizon Brilliant™ Blue family of dyes.   It is a polymer-based tandem dye developed exclusively by BD Biosciences.  With an excitation max of 485 nm and an emission max of 693 nm, BD Horizon BB700 can be excited by the 488 nm laser and detected in a standard PerCP-Cy™5.5 set (eg, 695/40-nm filter). This dye provides a much brighter alternative to PerCP-Cy5.5 with less cross laser excitation off the 405 nm and 355 nm lasers.

742120 Rev. 1
Format Details
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BB700
The BD Horizon Brilliant™ Blue 700 (BB700) Dye is part of the BD Horizon Brilliant™ Blue family of dyes. This tandem fluorochrome is comprised of a polymer-technology dye donor with an excitation maximum (Ex Max) of 476-nm and an acceptor dye with an emission maximum (Em Max) at 695-nm. Driven by BD innovation, BB700 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 695-nm (e.g., a 695/20-nm bandpass filter). The donor dye can be excited by the Violet (405 nm) laser and the acceptor dye can be excited by the red (627–640 nm) laser resulting in cross-laser excitation and fluorescence spillover. BB700 Reagents are significantly brighter than equivalent PerCP or PerCP-Cy5.5 reagents and are less sensitive to photobleaching. In addition, BB700 shows much less excitation by the violet (407-nm) laser resulting in less spillover. BB700 has minimal yellow green (562-nm) excitation and is ideal for instruments with both blue (488-nm) and yellow green (562-nm) lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
BB700
Blue 488 nm
476 nm
695 nm
742120 Rev.1
Citations & References
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Development References (14)

  1. Bluestone JA. New perspectives of CD28-B7-mediated T cell costimulation. Immunity. 1995; 2(6):555-559. (Clone-specific). 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
742120 Rev. 1

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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.