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: The IT2.2 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This IT2.2 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.2 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.

NovaFluor dyes are not compatible with DNA intercalating viability dyes. Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.

Each NovaFluor conjugate or kit is shipped with CellBlox Blocking Buffer. Use this buffer whenever staining with NovaFluor conjugates, including single-color compensation controls using cells. Whenever possible, we recommend adding CellBlox Blocking Buffer to antibody cocktails/master mixes prior to combining with cells. Add 5 µL per sample (regardless of the number of NovaFluors in your panel) to use the antibody cocktail as intended. For single-color controls, use 5 µL of CellBlox Blocking Buffer per 100µL of cell sample containing 10^3 to 10^8 cells.

NovaFluor conjugates are based on Phiton technology utilizing novel nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more

Excitation: 552 nm; Emission: 755 nm; Laser: 561 nm (Yellow) Laser