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Your Position: Home > Kits > CD28 > EP-150

CD28 : B7-1 [Biotinylated] Inhibitor Screening ELISA Kit

For research use only.

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IDComponentsSize
EP150-C01High-bind Plate1 plate
EP150-C02Human CD2820 μg
EP150-C03Biotinylated Human B7-150 μg
EP150-C04Anti-CD28 Neutralizing Antibody20 μg
EP150-C05Streptavidin-HRP5 μg
EP150-C06Coating Buffer12 mL
EP150-C0710xWashing Buffer 50 mL
EP150-C08Blocking Buffer50 mL
EP150-C09Substrate Solution12 mL
EP150-C10Stop Solution7 mL
  • 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.
  • Application

    This kit is developed for screening for inhibitors of human CD28 binding to human B7-1.

    It is for research use only.

  • Reconstitution
    Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.
  • 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.

  • 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 B7-1 to immobilized human CD28 in a functional ELISA assay, and employs a simple colorimetric ELISA platform. Briefly, we provide you with a human Biotinylated B7-1 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 B7-1-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:B7-1 binding will be determined by comparing OD readings among different experimental groups.

Typical Data Please refer to DS document for The assay protocol.
 CD28 TYPICAL DATA

INHIBITION OF CD28:B7-1[Biotinylated] BINDING BY ANTI-CD28 NEUTRALIZING ANTIBODY
Serial dilutions of Anti-CD28 Neutralizing antibody (Catalog # EP150-C04) (1:1 serial dilution, from 5 μg/mL to 0.01953125μg/mL) was added into CD28: Biotinylated 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 (QC tested).

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