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Human IL6/IL6R binding kit HTRF®

The Human IL6/IL6R binding kit is designed to identify human IL6/IL6R inhibitors.

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  • No-wash No-wash
  • All inclusive kit All inclusive kit
  • Validated with reference Ab Validated with reference Ab

The Human IL6/IL6R binding kit is designed to identify human IL6/IL6R inhibitors.

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Overview

IL6 is an inflammatory cytokine involved in various biological processes, including autoimmune diseases and cancers. It is produced by many cell lineages, including stromal cells, hematopoietic cells, epithelial cells, or muscle cells. IL6 binding to the membrane receptor IL6R induces receptor dimerization, and the activation of receptor-associated kinases (JAK1, JAK2, and Tyk2) which phosphorylate the receptor, leading to pathway stimulation. Anti-IL6 therapies have been developed for the treatment of various rheumatic diseases, as well as cytokine release syndrome.

Benefits

  • AUTOIMMUNE DISEASE STUDIES
  • IL6-BLOCKING DRUG DISCOVERY
  • KD DETERMINATION

Assay principle

The HTRF IL6/IL6 Binding kit is designed to measure the interaction between IL6 and IL6R proteins. Without treatment, IL6 binds to IL6R, and the binding of each detection antibody to its respective tagged protein generates an HTRF signal. In the presence of a compound or an anti-human blocking antibody, the IL6/IL6R interaction is disrupted and the HTRF signal decreases.

HTRF human IL6/IL6R binding kit assay principal

Assay Protocol

The Human IL6/IL6R binding kit can be run in 96- or 384-well low volume white plates (20 µL final). As described here, samples or standards are dispensed directly into the assay plate. The Tag1-IL6 protein and Tag2-IL6R proteins are then added, followed by the dispensing of the HTRF reagents: the anti Tag1 antibody labeled with europium cryptate and  the anti Tag2 antibody labeled with XL665. The reagents labeled 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.

HTRF human IL6/IL6R binding kit assay protocol

Inhibitory effects of IL6/IL6R blockers

The inhibitory effects of unlabeled IL6, IL6R, Tocilizumab (anti IL6R), and Sarilumab (anti IL6) were tested. No inhibition was observed using an anti CD20 antibody (Rituxiamb), confirming the specificity of the kit for the study of IL6 / IL6R interactions.

Validation of inhibitory effects of IL6/IL6R blockers

Best practices for pharmacological characterization of PPI inhibitors

Easy pharmacological characterization of PPI modulators. - Technical Notes

HTRF assays handle low- to high affinity protein-protein interactions

Deciphering low- and high affinity interactions - Application Notes

Nuclear receptor ligand identification with HTRF

Monitoring nuclear receptor binding with HTRF assays - Application Notes

HTRF addresses large protein-protein interaction complexes

Challenge large complexes with HTRF assays - Application Notes

HTRF PPI your dream assay served on a plate

Sandwiches aren't just for eating - Infographics

A brief history of Protein-Protein Interactions

How well do you know PPI? - Infographics

HTRF Product Catalog 2020 July update

All your HTRF assays in one document! - Catalog

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 - Application Notes

Product Insert IL6/IL6R (h) Kit / 64BDIL6PEG-64BDIL6PEH

64BDIL6PEG-64BDIL6PEH - Product Insert

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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