Invitrogen

CD278 (ICOS) Monoclonal Antibody (ISA-3), Functional Grade, eBioscience™

Product Image Gallery

CD278 (ICOS) Antibody in Flow Cytometry (Flow) — Staining of Anti-Human CD3/CD28-stimulated (Product # 16-0039-81 and Product # 16-0289-81) with Anti-Human CD278 (ICOS) FITC (left) and PE (right). Appropriate isotype controls were used (open histogram). Total viable cells were used for analysis.

CD278 (ICOS) Antibody in T-Cell Activation (TCA) — Figure 6. ICOS is upregulated in Melan-A-specific CD8 + T cells isolated after combined therapy and is involved in chemoimmunotherapy-induced AKT activation. (A) Top panel shows representative staining with anti-ICOS (open histogram) or isotype (filled histogram) mAbs, in two T-cell clones isolated from Arm1 (Pt08.90) and Arm2 (Pt15.26) patients after the treatments (POST). Bottom panel shows pooled data of ICOS expression in Melan-A-specific T-cell clones (n = 15 from two patients of Arm1, n = 18 from three patients of Arm2), isolated before (PRE) and after therapy (POST). Cells were stained after 48 h stimulation with plate-bound anti-CD3 mAb (2 ug/mL) plus rIL-2 and shown in both scatter-plot graph and box-and-whisker diagram, with 5-95 percentile. Each dot represents the mean value out of three-five independent experiments. The mean +- SEM of each group is shown. (B) Correlation between p-AKT and ICOS expression. T-cell clones isolated after both treatments were grouped according to pSerAKT expression levels and the percentage of ICOS + cells (bottom panel) and shows higher expression of ICOS in AKT high T cells. Top panel, percentage of clones with ICOS levels > 30% cut-off in p-AKT neg/low versus p-AKT high groups. p value was calculated with one-tail Fisher's exact test. * / p <= 0.05, ** / p <= 0.01, *** / p <= 0.001, Mann-Whitney two-sample test (*) and two-tail Student's test ( ), respectively. (C) Left panel: effect of ICOS blockade on AKT activation (p-Ser473-AKT)

Product Details

16-9948-82

Applications	Tested Dilution	Publications
Flow Cytometry (Flow)	0.5 µg/test	View 8 publications 8 publications
Functional Assay (FN)	Assay-Dependent	View 2 publications 2 publications
T-Cell Activation (TCA)	-	View 2 publications 2 publications

Product Specifications
Species Reactivity	Human
Published species	Human
Host/Isotype	Mouse / IgG1, kappa
Recommended Isotype Control	Mouse IgG1 kappa Isotype Control (P3.6.2.8.1), Functional Grade, eBioscience™
Class	Monoclonal
Type	Antibody
Clone	ISA-3
Conjugate	Functional Grade Alexa Fluor 700 APC APC-eFluor 780 Biotin Brilliant UV 661 eFluor 450 FITC NovaFluor Red 660 NovaFluor Red 710 NovaFluor Yellow 700 NovaFluor Yellow 730 PE PE-Cyanine5.5 PE-Cyanine7 PE-eFluor 610 PerCP-eFluor 710 Super Bright 436 Super Bright 600 Super Bright 645 Super Bright 780 Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Affinity chromatography
Storage buffer	PBS, pH 7.2
Contains	no preservative
Storage conditions	4° C
RRID	AB_469295

Product Specific Information

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 # 14-9949-82), 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 13-9948-82 or 12-9948-42 rather than 11-9948-42.

Applications Reported: The ISA-3 antibody has been reported for use in flow cytometric analysis. It has also been reported in in vitro functional assays.

Applications Tested: The ISA-3 antibody has been tested by flow cytometric analysis of unstimulated and PHA-activated (2 days) human blood cells. This can be used at less than or equal to 0.5 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

Storage and handling: Use in a sterile environment.

Filtration: 0.2 µm post-manufacturing filtered.

Purity: Greater than 90%, as determined by SDS-PAGE.

Endotoxin Level: Less than 0.001 ng/µg antibody, as determined by LAL assay.

Aggregation: Less than 10%, as determined by HPLC.

Target Information

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.

Product References

Impaired HA-specific T follicular helper cell and antibody responses to influenza vaccination are linked to inflammation in humans.

eLife

Hill DL,Whyte CE,Innocentin S,Lee JL,Dooley J,Wang J,James EA,Lee JC,Kwok WW,Zand MS,Liston A,Carr EJ,Linterman MA