Human CD47/SIRP alpha biochemical binding kit
CD47 / SIRP alpha biochemical binding assay
Human PD1/PD-L1 biochemical binding assay HTRF®
This immune checkpoint assay is designed to identify human PD1/PD-L1 checkpoint inhibitors.
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All inclusive kit
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Scalable from 96 to 1536 format
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Validated with reference Ab
Overview
Programmed cell death protein 1 (PD1) is an immune checkpoint receptor that regulates T cell response. Its ligand, programmed death-ligand 1 (PD-L1), is commonly over-expressed on a tumor cell surface. When PD1 is bound to PD-L1, T cell response is suppressed, contributing to tumor immune resistance. Checkpoint inhibitors blocking PD1/PD-L1 complex formation are generating considerable interest in cancer immunotherapy.
Benefits
- T CELL RESPONSE
- IMMUNOTHERAPY TO FIGHT CANCER
- DISCOVER IMMUNE CHECKPOINT BLOCKING DRUGS
Assay principle
The PD1/PD-L1 binding assay includes tagged human recombinant immune checkpoint partners (PD1 and PD-L1) and labelled anti-tag reagents for HTRF detection. Without an inhibitor, PD1 binds to PD-L1, and the binding of each detection reagent to its tagged target generates an HTRF signal. In the presence of compound, standard, or an antibody blocking the PD1 /PD-L1 interaction, the HTRF signal decreases.
Assay protocol
This PD1/PD-L1 binding assay can be run in a 96- or 384-well low volume white plate (20 µL final). As described here, samples or standards are dispensed directly into the assay plate, and the tagged PD1 & PD-L1 proteins are then added, followed by the dispensing of the HTRF reagents. The reagents labelled with HTRF fluorophores may be pre-mixed and added in a single dispensing step. No washing steps are needed. The protocol can be further miniaturized or upscaled by simply resizing each addition volume proportionally.
Inhibitory effect of blocking antibodies and small molecules
The inhibitory effects of three antibodies and two small molecules known to inhibit the PD1 / PD-L1 interaction were tested. The two anti-PD1 antibodies (pembrolizumab & nivolumab) were shown to inhibit the interaction with IC50 values of 2.0 & 2.4 nM respectively. The anti-PD-L1 antibody (atezolizumab) disrupted the interaction, with an IC50 of 3.9 nM. Two small molecule PD1 / PD-L1 inhibitors 1 & 3 show IC50s of 177 & 146 nM in these assay conditions.
Inhibitory effect of the PD1 / PD-L1 standard
The PD1 / PD-L1 standard stock solution provided in the kit was diluted to prepare the 8-point dose response curve using 5-fold serial dilutions. As described in the assay protocol, standards were dispensed into the assay plate, and the tagged PD1 & PD-L1 proteins were then added, followed by the dispensing of the HTRF reagents. The data presented in the graph show that the PD1 / PD-L1 interaction is inhibited, with an IC50 of 3.8 nM.
Best practices for pharmacological characterization of PPI inhibitors
Easy pharmacological characterization of PPI modulators. - Notes techniques
HTRF assays handle low- to high affinity protein-protein interactions
Deciphering low- and high affinity interactions - Notes d'application
Nuclear receptor ligand identification with HTRF
Monitoring nuclear receptor binding with HTRF assays - Notes d'application
HTRF addresses large protein-protein interaction complexes
Challenge large complexes with HTRF assays - Notes d'application
HTRF PPI your dream assay served on a plate
Sandwiches aren't just for eating - Infographies
A brief history of Protein-Protein Interactions
How well do you know PPI? - Infographies
All your HTRF assays in one document! - Catalogue
A guide to Homogeneous Time Resolved Fluorescence
General principles of HTRF - Guides
How HTRF compares to Western Blot and ELISA
Get the brochure about technology comparison. - Brochures
Virology research solutions using HTRF Protein-Protein Interaction assays
See how peer researchers challenge the viral life cycle with PPI assays - Notes d'application
Product Insert PD1/PD-L1 Kit2 / 64PD1PEG-64PD1PEH
64PD1PEG-64PD1PEH - Protocoles
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Complete autoimmunity review in a single White Paper - Guides
Autoimmune diseases and therapeutic strategies
Check out our new guide featuring valuable insights on autoimmunity - Guides
Safety Data Sheet (DEU) PD1/PD-L1 Kit2 / 64PD1PEG
64PD1PEG - Fiches de sécurité
Safety Data Sheet (ELL) PD1/PD-L1 Kit2 / 64PD1PEG
64PD1PEG - Fiches de sécurité
Safety Data Sheet (FRA-FR) PD1/PD-L1 Kit2 / 64PD1PEG
64PD1PEG - Fiches de sécurité
Safety Data Sheet (ITA) PD1/PD-L1 Kit2 / 64PD1PEG
64PD1PEG - Fiches de sécurité
Safety Data Sheet (SPA) PD1/PD-L1 Kit2 / 64PD1PEG
64PD1PEG - Fiches de sécurité
Safety Data Sheet (ENG-GB) PD1/PD-L1 Kit2 / 64PD1PEG
64PD1PEG - Fiches de sécurité
Safety Data Sheet (ENG-US) PD1/PD-L1 Kit2 / 64PD1PEG
64PD1PEG - Fiches de sécurité
Safety Data Sheet (DEU) PD1/PD-L1 Kit2 / 64PD1PEH
64PD1PEH - Fiches de sécurité
Safety Data Sheet (ELL) PD1/PD-L1 Kit2 / 64PD1PEH
64PD1PEH - Fiches de sécurité
Safety Data Sheet (FRA-FR) PD1/PD-L1 Kit2 / 64PD1PEH
64PD1PEH - Fiches de sécurité
Safety Data Sheet (ITA) PD1/PD-L1 Kit2 / 64PD1PEH
64PD1PEH - Fiches de sécurité
Safety Data Sheet (SPA) PD1/PD-L1 Kit2 / 64PD1PEH
64PD1PEH - Fiches de sécurité
Safety Data Sheet (ENG-GB) PD1/PD-L1 Kit2 / 64PD1PEH
64PD1PEH - Fiches de sécurité
Safety Data Sheet (ENG-US) PD1/PD-L1 Kit2 / 64PD1PEH
64PD1PEH - Fiches de sécurité
Plate Reader Requirement
Choosing the right microplate reader ensures you’ll get an optimal readout. Discover our high performance reader, or verify if your lab equipment is going to be compatible with this detection technology.
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