ISK-010

CTLA4; CD80

ID Components Size (96 tests)
ISK-010-1 High-bind Plate 1 plate
ISK-010-2 Human B7-1 20 μg
ISK-010-3 Biotinylated Human CTLA-4 10 μg
ISK-010-4 Anti-CTLA-4 Neutralizing Antibody 20 μg
ISK-010-5 Streptavidin-HRP 10 μg
ISK-010-6 Coating Buffer 12 mL
ISK-010-7 10xWashing Buffer 50 mL
ISK-010-8 Blocking Buffer 50 mL
ISK-010-9 Substrate Solution 12 mL
ISK-010-10 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

10-0.005μg/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 CTLA-4 to immobilized human B7-1 in a functional ELISA assay, and employs a simple colorimetric ELISA platform. Briefly, we provide you with a human Biotinlated CTLA-4 protein, a human B7-1 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 B7-1.
b) Add your molecule of interest to the tests.
c) Add human CTLA-4-Biotin to bind the coated human B7-1.
d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.
Finally, the half maximal inhibitory concentration (IC50) of your compound to CTLA-4: B7-1 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 0.005μg/mL) was added into B7-1: Biotinlated CTLA-4 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.