|Flow Cytometry (Flow)||5 µL (0.125 µg)/test|
|Flow Cytometry (Flow)||See 2 publications below|
|Functional Assay (FN)||See 1 publications below|
|Immunofluorescence (IF)||See 1 publications below|
|Miscellaneous PubMed (MISC)||See 1 publications below|
|Tested Species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG1, kappa|
|Recommended Isotype Control||Mouse IgG1 kappa Isotype Control, PE-Cyanine7, eBioscience™|
|Excitation/Emission Profile||View spectra|
|Storage buffer||PBS, pH 7.2, with 0.1% gelatin, 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage conditions||4° C, store in dark, DO NOT FREEZE!|
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 # 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.
Applications Tested: This ISA-3 antibody has been pre-titrated and tested by flow cytometric analysis of stimulated normal human peripheral blood cells. This can 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.
Light sensitivity: This tandem dye is sensitive photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL cell sample + 100 µL IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 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: 488-561 nm; Emission: 775 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.
Filtration: 0.2 µm post-manufacturing filtered.
CD278 belongs to the CD28 and CTLA-4 cell-surface receptor family and is highly expressed in activated T-cells. It is essential both for efficient interaction between T and B-cells and for normal antibody responses to T-cell dependent antigens. It enhances all basic T-cell responses to a foreign antigen, namely proliferation, secretion of lymphokines, up-regulation of molecules that mediate cell-cell interaction, and effective help for antibody secretion by B-cells.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: Activation-inducible lymphocyte immunomediatory molecule; CD278; inducible costimulator; inducible T-cell co-stimulator; Inducible T-cell costimulator
Gene Aliases: AILIM; CD278; CVID1; ICOS
UniProt ID: (Human) Q9Y6W8
Entrez Gene ID: (Human) 29851
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