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-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 μL (0.25 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. Super Bright 645 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 645 nm. We recommend using a 660/20 bandpass filter.Please make sure that your instrument is capable of detecting this fluorochrome. When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Staining Buffer (Product No. SB-4400) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining 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 No. 00-8222) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product No. 00-5333) for up to 3 days in the dark at 4 degrees 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. Excitation: 405 nm; Emission: 645 nm; Laser: Violet Laser CD86 is one of two ligands (the other CD80) for CTLA4 and CD28. CD86 acts as costimulatory molecule in eliciting T-cell help during antigen presentation. Antigen presentation in the absence of sufficient co-stimulation involving CD86/CD80 can induce tolerance. CD80 appears to play a role distinct from CD80 in T helper cell differentiation. CD86 is a type I membrane protein that is a member of the immunoglobulin superfamily. The CD86 protein is expressed by antigen-presenting cells, and it is the ligand for two proteins at the cell surface of T cells, CD28 antigen and cytotoxic T-lymphocyte-associated protein 4. Binding of CD86 with CD28 antigen is a costimulatory signal for activation of the T-cell. Binding of CD86 with cytotoxic T-lymphocyte-associated protein 4 negatively regulates T-cell activation and diminishes the immune response. Alternative splicing results in two transcript variants encoding different isoforms of CD86. Additional transcript variants have been described for CD86, but their full-length sequences have not been determined. Diseases associated with CD86 dysfunction include gallbladder squamous cell carcinoma and myocarditis.
|4° C, store in dark, DO NOT FREEZE!|
|Super Bright 645|
|PBS with BSA and 0.09% sodium azide; pH 7.2|