Product Information

Contents: Affinity Purified anti-human ICOS (CD278)
Catalog Number: 14-9948
Sizes: 25 ug, 100 ug
Formulation: Phosphate buffer pH 7.2,
150 mM NaCl, 0.09% NaN₃
Storage Conditions: Store at 4°C.
Avoid repeated freeze/thaw cycles.
Clone: ISA-3
Isotype: Mouse IgG1, κ

Surface staining of CD3 + CD28 stimulated human PBMC with anti-human ICOS (ISA-3) FITC (left), and PE (right). Appropriate isotype controls were used (open histogram). Total viable cells were used for analysis

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
14-9948	Affinity Purified anti-human ICOS (CD278)	N/A	N/A	FA FC IH/F IP
16-9948	Functional Grade* Purified anti-human ICOS (CD278)	N/A	N/A	FA FC
13-9948	Biotin anti-human ICOS (CD278)	N/A	N/A	FC
11-9948	FITC anti-human ICOS (CD278)	488	518	FC
12-9948	Phycoerythrin (PE) anti-human ICOS (CD278)	488	575	FC
25-9948	Phycoerythrin-Cy7 (PE-Cy7) anti-human ICOS (CD278)	488	760	FC
*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 Pacific Blue® and APC-Alexa Fluor® 750 conjugated products with eFluor™ 450 and APC-eFluor™ 780 conjugated products, respectively.

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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 (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

For research use only, not for diagnostic or therapeutic use. The ISA-3 antibody has been reported for use in flow cytometric analysis, immunoprecipitation, and immunohistochemical staining of frozen tissue sections. It has also been reported in in vitro functional assays. (Please use Functional Grade purified ISA-3, cat. 16-9948, in functional assays.)

Applications Tested

The ISA-3 antibody has been tested by flow cytometric analysis of unstimulated and PHA-activated (2 days) human blood leukocytes. This can be used at less than or equal to 0.5 μg per million cells in a 100 μl total staining volume. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

References

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.

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.

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.

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.

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