PD-L1 Inhibitor Screening ELISA Kit

Catalog No. ABIN2762507

Product Details

Target: PD-L1 (CD274 (PD-L1))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.0125 μg/mL - 0.2 μg/mL

Minimum Detection Limit: 0.0125 μg/mL

Application: ELISA, Screening Assay (ScA)

Purpose: PD-L1 Inhibitor Screening ELISA Kit: This kit is developed for screening for inhibitors of human PD-1 binding to human PD-L1.

Components: High-bind Plate
Human PD-L1
Anti-PD-1 Neutralizing Antibody
Human PD-1-Biotin
Streptavidine-HRP
Coating Buffer
10x Washing Buffer
Blockung Buffer
Substrate Solution
Stop Solution

Application Details

Application Notes: The antibody pair is useful for screening for inhibitors of human PD-1 binding to human PD-L1.

Comment:

METHOD VERIFICATION
PD-1 [BIOTINYLATED]:PD-L1 BINDING IN THE ABSENCE OF INHIBITORS:
Immobilized human PD-L1 protein at 2 μg/mL (100 μL/well) can bind human PD-1-Biotin with a linear range of 0.01875-0.3 μg/mL when detected by Streptavidin-HRP. Background was subtracted from data points before curve fitting.

INHIBITION OF PD-1 [BIOTINYLATED] : PD-L1 BINDING BY ANTI-PD-1 NEUTRALIZING ANTIBODY
Serial dilutions of anti-PD-1 neutralizing antibody (1:2 serial dilutions, from 10 μg/mL to 0.078 μg/mL) was added into PD-L1 : PD-1-Biotin binding reactions. The assay was performed according to the above described protocol. Background was subtracted from data points prior to log transformation and curve fitting.
Preparation of Standard Serial Dilutions
1) Pipette 480 μL Dilution Buffer into tube Std-1 and 150 μL Dilution Buffer into the remaining tubes (tube Std.-2 to tube Std.-8).
2) Add 20 μL anti-PD-1 neutralizing antibody stock solution (250 μg/mL) to tube Std.-1 to make the final concentration 10 μg/mL. Mix well.
3) To make a 1:2 serial dilutions, pipette 150 μL solution to the next tube as illustrated in Fig.3.
IMPORTANT: Mix thoroughly at each step during the dilution process.

Protocol: This assay employs a simple colorimetric ELISA platform, which measures the binding between immobilized human PD-L1 and in-house developed biotinylated PD-1 protein. This product is uniquely suitable for rapid high-throughput screening of putative PD-1 and PD-L1 inhibitors. Briefly, we provide you with a human PD-1-Biotin protein, a human PD-L1 protein, an anti-PD-1 neutralizing antibody (as method verified Std.), and Streptavidin-HRP reagent. Your experiment will include 4 simple steps:

• Coat the plate with human PD-L1.
• Add your molecule of interest to the tests.
• Add human PD-1-Biotin to bind the coated human PD-L1.
• Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.

Finally, the ability of your compound to inhibit PD-1 : PD-L1 binding will be determined by comparing OD readings among different experimental groups

Restrictions: For Research Use only

Handling

Storage: 4 °C

Storage Comment: The unopened kit is stable for 12 months from the date of manufacture if stored at 2°C to 8°C.

Target Details

Target: PD-L1 (CD274 (PD-L1))

Alternative Name: PD-1,PD-L1 (PD-L1 Products)

Synonyms: B7-H Kit, B7H1 Kit, PD-L1 Kit, PDCD1L1 Kit, PDCD1LG1 Kit, PDL1 Kit, A530045L16Rik Kit, B7h1 Kit, Pdcd1l1 Kit, Pdcd1lg1 Kit, Pdl1 Kit, RGD1566211 Kit, CD274 molecule Kit, CD274 antigen Kit, programmed cell death 1 ligand 2 Kit, CD274 Kit, Cd274 Kit, PDCD1LG2 Kit

Background: Immune checkpoint pathway is a focal point of today's cancer research. PD-1 is one of the best characterized checkpoint proteins. The binding between PD-1 and its ligand PD-L1 suppresses T-cell activation and allows cancer cells to escape from body's immune surveillance. Therefore, the pharmaceutical inhibition of PD-1 or its ligand has been considered a promising strategy by many oncologists.