CD86 (B7-2) Monoclonal Antibody (GL1), Brilliant Violet™ 786, eBioscience™

Product Specific Information

Description
The GL1 monoclonal antibody reacts with mouse CD86, an ~80 kDa surface receptor also known as B7-2. CD86 and CD80 are members of the B7 family of costimulatory molecules. CD86 is expressed at low level on B cells, macrophages, and dendritic cells and is upregulated on B cells through a variety of surface stimuli including the BCR complex, CD40 and some cytokine receptors. CD86 is also expressed by activated mouse T cells and thioglycolate-elicited peritoneal cells. In addition to CD80 (B7-1), CD86 is a counter-receptor for the T cell surface molecules CD28 and CD152 (CTLA-4). This interaction plays a critical role in T-B crosstalk, T cell costimulation, autoantibody production and Th2-mediated Ig production. The kinetics of upregulation of CD86 upon stimulation, supports its major contribution during the primary phase of an immune response.

Applications Tested
This GL1 antibody has been tested by flow cytometric analysis of mouse splenocytes. This may be used at less than or equal to 0.5 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

Blocking Buffers
When using two or more Super Bright, Brilliant Violet™, Brilliant Ultra Violet™, or other polymer dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) or Brilliant Stain Buffer (Product # 00-4409-75) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer or Brilliant Stain Buffer for more information.

Light sensitivity
This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.

Fixation
• Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation.
• Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
• Our internal testing suggests that Brilliant Violet™ 786 (BV786) is not compatible with methanol-based fixation.

Excitation: 407 nm; Emission: 786 nm; Laser: Violet Laser.

BRILLIANT ULTRA VIOLET™ is a trademark or registered trademark of Becton, Dickinson and Company or its affiliates, and is used under license. Powered by Sirigen™.

Target Information

CD86, along with CD80, is a member of the B7 family of costimulatory molecules and plays a crucial role in T cell activation and immune response regulation. CD86 is expressed at low levels on B cells, macrophages, and dendritic cells, and its expression is upregulated on B cells through various stimuli, including the BCR complex, CD40, and certain cytokine receptors. As a type I membrane protein and member of the immunoglobulin superfamily, CD86 serves as a ligand for the T cell surface proteins CD28 and CTLA-4 (CD152). The interaction between CD86 and CD28 provides a costimulatory signal essential for T cell activation during antigen presentation, while binding with CTLA-4 negatively regulates T cell activation, diminishing the immune response. This interaction is critical for T-B cell crosstalk, T cell costimulation, autoantibody production, and Th2-mediated Ig production. The kinetics of CD86 upregulation upon stimulation suggest its significant contribution during the primary phase of an immune response. CD86 and CD80 have distinct roles in T helper cell differentiation, and insufficient co-stimulation involving these molecules can induce tolerance. Alternative splicing of CD86 results in two transcript variants encoding different isoforms, with additional variants described but not fully sequenced. Dysfunction in CD86 is associated with diseases such as gallbladder squamous cell carcinoma and myocarditis.