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 Reported: This GL1 antibody has been reported for use in flow cytometric analysis.

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.

Brilliant Violet™ 786 (BV786) is a tandem dye that emits at 786 nm and is intended for use on cytometers equipped with a violet (405 nm) laser. Please make sure that your instrument is capable of detecting this fluorochrome.

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-4401-42) 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 # 00-8222-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-54) 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.

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