Staining of normal human peripheral blood cells either unstimulated (blue histogram) or stimulated for 3 days with anti-CD3/CD28 (purple histogram) with Anti-Human CD278 (ICOS) PE-Cy7. Total viable cells were used for analysis.
Contents: Anti-Human CD278 (ICOS) PE-Cy7 Catalog Number: 25-9948 Concentration: 5 μl (0.125 μg)/test Formulation: aqueous buffer, 0.09% sodium azide, may contain carrier protein/stabilizer Storage Conditions: Store at 2-8°C. Do not freeze. Light sensitive material. This Cy7 tandem dye is sensitive to photo-induced oxidation. Protect this vial from light during storage, handling & experimental procedures. Clone: ISA-3 Host/Isotype: Mouse IgG1, κ
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25-9948-41
25 tests
25-9948-42
100 tests
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*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µL to 5 µL. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.
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.
This tandem dye is sensitive to photo-induced oxidation. Protect this reagent from light during storage and all handling steps. When staining cells with PE-Cy7-conjugated antibodies, always protect samples from light.
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)/test 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 105 to 108 cells/test.
References
Quiroga MF, Pasquinelli V, Martínez GJ, Jurado JO, Zorrilla LC, Musella RM, Abbate E, Sieling PA, García VE.Inducible costimulator: a modulator of IFN-gamma production in human tuberculosis. J Immunol. 2006 May 15;176(10):5965-74.(ISA-3, FA, PubMed)
Grimbacher B, Hutloff A, Schlesier M, Glocker E, Warnatz K, Drager R, Eibel H, Fischer B, Schaffer AA, Mages HW, Kroczek RA, Peter HH. 2003. Homozygous loss of ICOS is associated with adult-onset common variable immunodeficiency. Nat Immunol. 4(3):261-8.
Buonfiglio D, Bragardo M, Redoglia V, Vaschetto R, Bottarel F, Bonissoni S, Bensi T, Mezzatesta C, Janeway Jr CA, Dianzani U. 2000. The T cell activation molecule H4 and the CD28-like molecule ICOS are identical. Eur J Immunol. 30(12):3463-7.
Beier KC, Hutloff A, Dittrich AM, Heuck C, Rauch A, Buchner K, Ludewig B, Ochs HD, Mages HW, Kroczek RA. 2000. Induction, binding specificity and function of human ICOS. Eur. J. Immunol. 30, 3707.
Hutloff A, Dittrich AM, Beier KC, Eljaschewitsch B, Kraft R, Anagnostopoulos I, Kroczek RA. 1999. ICOS is an inducible T-cell co-stimulator structurally and functionally related to CD28. Nature. 397(6716):263-6.
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