ISK-008

CD80; CTLA4

ID Components Size (96 tests)
ISK-008-1 High-bind Plate 1 plate
ISK-008-2 Human CTLA-4 10 μg
ISK-008-3 Biotinylated Human B7-1 10 μg
ISK-008-4 Anti-CTLA4 Neutralizing Antibody 20 μg
ISK-008-5 Streptavidin-HRP 10 μg
ISK-008-6 Coating Buffer 12 mL
ISK-008-7 10xWashing Buffer 50 mL
ISK-008-8 Blocking Buffer 50 mL
ISK-008-9 Substrate Solution 12 mL
ISK-008-10 Stop Solution 7 mL

Cytotoxic T lymphocyte associated ANTIgen-4 (CTLA-4), also known as CD152, is a kind of leukocyte differentiation antigen and a transmembrane receptor on T cells, sharing B7 ligand with CD28. The binding of CTLA-4 with B7 induces T cells to be non-reactive and participates in the negative regulation of immune response. The recombinant CTLA-4 Ig can effectively and specifically inhibit cellular and humoral immune responses in vivo and in vitro, and has significant therapeutic effects on transplant rejection and various autoimmune diseases with very low toxicity. It is considered as a promising new immunosuppressive drug at present.

96 tests

10-0.078μg/mL

This inhibitor screening ELISA kit is designed to facilitate the identification and characterization of new CTLA-4 pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated human B7-1 to immobilized human CTLA-4 in a functional ELISA assay, and employs a simple colorimetric ELISA platform. Briefly, we provide you with a human Biotinlated B7-1 protein, a human CTLA-4 protein, an anti-CTLA-4 neutralizing antibody (as method verified Std.), and Streptavidin-HRP reagent.
Your experiment will include 4 simple steps:
a) Coat the plate with human CTLA-4.
b) Add your molecule of interest to the tests.
c) Add human B7-1-Biotin to bind the coated human CTLA-4.
d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.
Finally, the half maximal inhibitory concentration (IC50) of your compound to B7-1: CTLA-4 binding will be determined by comparing OD readings among different experimental groups.

This kit is developed for screening for inhibitors of human CTLA-4 binding to human B7-1

See Certificate of Analysis for details of reconstitution instruction and specific concentration.

The unopened kit is stable for 12 months from the date of manufacture if stored at 2°C to 8°C.
The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.

Serial dilutions of Anti-CTLA-4 Neutralizing antibody(1:1 serial dilution, from 10 μg/mL to 0.078μg/mL) was added into CTLA-4: Biotinlated B7-1 binding reactions. The assay was performed according to the protocol described below. Background was subtracted from data points prior to log transformation and curve fitting.