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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 (Product # 14-9949-82) , 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 Product # 13-9948-82 or Product # 12-9948-42 rather than Product # 11-9948-42 .
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 (0.25 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.
PE-eFluor® 610 can be excited with laser lines from 488-561 nm and emits at 607 nm. We recommend using a 610/20 band pass filter (equivalent to PE-Texas Red®). Please make sure that your instrument is capable of detecting this fluorochome.
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.
Excitation: 488-561 nm; Emission: 607 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.
Filtration: 0.2 µm post-manufacturing filtered.
ICOS (CD278) encodes a protein that is a member of the CD28 and CTLA-4 cell-surface receptor family. It functions as a homodimer and has a crucial role in cell-cell signaling, immune responses, and the regulation of cell proliferation. ICOS has been linked to various disorders, including Immunodeficiency, Common Variable, 1 and Common Variable Immunodeficiency.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.
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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