IgD Monoclonal Antibody (IA6-2), Super Bright™ 780, eBioscience™, Invitrogen™

Description

Description: The ISA-3 monoclonal antibody reacts with human ICOS (Inducible COStimulatory molecule), also known as H4, CRP-1 and AILIM. ICOS is a T cell specific activation molecule and a third member of the CD28/CTLA-4 family. Human ICOS has a relative molecular mass of 55-60 kDa, composed of 27 kDa and 29 kDa chains. Human ICOS on activated T cells has potent costimulatory activity for T cell activation and is required for humoral immune responses, in particular for memory B cell and plasma cell generation. ICOS binds to its ligand, B7h/B7RP-1 expressed on activated APCs (antigen presenting cells) and on a number of inflamed peripheral tissues. Plate-bound ISA-3 is costimulatory for T cells and induces production of IL-4, IL-5, IL-10 and other cytokines, but not IL-2. ISA-3 has the same reactivity pattern and characteristics as F44. ISA-3 was generated against the human ICOS antigen. C398.4A, anti-mouse ICOS/H4 (catalog No. 14-9949), was shown to cross-react with human ICOS but binds to an epitope different from ISA-3. C398.4A stains activated cells brighter than ISA-3; however, it also exhibits higher staining of non-activated human peripheral blood or isolated PBMC. To achieve the brightest staining of ICOS on activated human T cells, please use 13-9948 or 12-9948 rather than 11-9948. Applications Reported: This ISA-3 antibody has been reported for use in flow cytometric analysis.

In some experiments, we have observed that compensation values for Super Bright 780-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres (Product No. 01-2222-42) as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads (Product No. A10497). 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°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: 780 nm; Laser: Violet Laser Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company. This type I transmembrane protein is co-expressed on the surface of mature naive B cells with membrane IgM. IgD also exists as a soluble form in blood serum. Studies have demonstrated that IgD associates with the B cell receptor and participates in mediating signal transduction upon antigen binding.