Description: The IT2.2 monoclonal antibody reacts with human 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 levels 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. In addition to CD80 (B7-1), CD86 is a counter-receptor for the T cell surface molecules CD28 and CD152 (CTLA-4). The interaction of CD86 with its ligands 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 IT2.2 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This IT2.2 antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 µL (0.125 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.

Brilliant Ultra Violet™ 737 (BUV737) is a tandem dye that emits at 732 nm and is intended for use on cytometers equipped with an ultraviolet (355 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) 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 Ultra Violet™ 737 (BUV737) is compatible with short-term methanol-based fixation, but should not be stored in buffers containing methanol for longer than one hour.

Excitation: 355 nm; Emission: 732 nm; Laser: Ultraviolet Laser.

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