ID | Components | Size |
EP151-C01 | High-bind Plate | 1 plate |
EP151-C02 | Human CD28 | 15 ug |
EP151-C03 | Biotinylated Human B7-2 | 50 ug |
EP151-C04 | Anti-CD28 Neutralizing Antibody | 20 ug |
EP151-C05 | Streptavidin-HRP | 5 ug |
EP151-C06 | Coating Buffer | 12 mL |
EP151-C07 | 10xWashing Buffer | 50 mL |
EP151-C08 | Blocking Buffer | 50 mL |
EP151-C09 | Substrate Solution | 12 mL |
EP151-C10 | Stop Solution | 7 mL |
背景(Background)
T-cell-specific surface glycoprotein CD28 is also known as TP44, is a single-pass type I membrane protein which contains one Ig-like V-type (immunoglobulin-like) domain. is one of the molecules expressed on T cells that provide co-stimulatory signals, which are required for T cell activation. CD28 is the receptor for B7-1 (CD80) and B7-2 (CD86). When activated by Toll-like receptor ligands, the CD80 expression is upregulated in antigen presenting cells (APCs). The CD86 expression on antigen presenting cells is constitutive. CD28 is the only B7 receptor constitutively expressed on naive T cells.
应用说明(Application)
This kit is developed for screening for inhibitors of human CD28 binding to human B7-2.
It is for research use only.
重构方法(Reconstitution)
Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.
存储(Storage)
Unopened kit should be stored at 2°C -8°C upon receiving. Find the expiration date on the outside packaging and do not use reagents past their expiration date.
The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.
原理(Assay Principles)
This inhibitor screening ELISA kit is designed to facilitate the identification and characterization of new CD28 pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated human B7-2 to immobilized human CD28 in a functional ELISA assay, and employs a simple colorimetric ELISA platform. Briefly, we provide you with a human Biotinylated B7-2 protein, a human CD28 protein, an anti-CD28 neutralizing antibody (as method verified Std.), and Streptavidin-HRP reagent.
Your experiment will include 4 simple steps:
a) Coat the plate with human CD28.
b) Add your molecule of interest to the tests.
c) Add human B7-2-Biotin to bind the coated CD28.
d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.
Finally, the half maximal inhibitory concentration (IC50) of your compound to CD28:B7-2 binding will be determined by comparing OD readings among different experimental groups.
典型数据-Typical Data Please refer to Ds document for the assay protocol.
INHIBITION OF CD28:B7-2[Biotinylated] BINDING BY ANTI-CD28 NEUTRALIZING ANTIBODY
Serial dilutions of Anti-CD28 Neutralizing antibody (Catalog # EP151-C04) (1:1 serial dilution, from 5 μg/mL to 0.01953125μg/mL) was added into CD28: Biotinylated B7-2 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 (QC tested).