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Product Details
FAB
- Comprehensive validation - Validated with various antibody drugs.
- Simple and fast operation - No complicated washing steps, significantly reducing time.
- High batch consistency - Strict control over raw materials and finished product quality, ensuring a stable supply.
- Accurate and reliable results - High sensitivity with minimal matrix effects.
- High throughput capability - Support multiple product specifications, ideal for high-throughput screening.
- Fast completion - Results in just 1 hour.
Product Specifications
Assay TypeCompetition-TR-FRETReactivityHumanRegulatory StatusRUOAssay Time1 hrSample volume10 μLProduct Overview
The Human IL-6 / IL-6 R alpha Inhibitor Screening Kit (TR-FRET) is based on a homogeneous (no wash) competition TR-FRET technology (Time-Resolved Fluorescence Resonance Energy Transfer) to screen for inhibitors of human IL-6 binding to IL-6 R alpha within one hour. It can also be used as a universal detection tool to identify the ability of human IL-6 to bind to human IL-6 R alpha.
Storage
1. Unopened kit should be stored at 2℃-8℃ upon receiving.
2. Find the expiration date on the outside packaging and do not use reagents past their expiration date.
3. The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.
Materials Provided
IDComponentsSizeFRTP023-C01Human IL-6 Protein Europium-chelate100 tests/500 testsFRTP023-C02FA Labeled Human IL-6 R alpha100 tests/500 testsFRTP023-C03Anti- IL-6 Neutralizing Antibody20 μg/100 tests 100 μg/500 testsDB-04TR-FRET Sample Dilution Buffer, pH7.450 mL/100 tests & 500 testsDB-05TR-FRET Detection Buffer, pH7.450 mL/100 tests & 500 testsAssay Principles
This Human IL-6 / IL-6 R alpha Inhibitor Screening Kit (TR-FRET) is based on TR-FRET technology (Time-Resolved Fluorescence Resonance Energy Transfer). This assay uses the mixture of biotinylated human IL-6 and Europium-chelate labeled streptavidin as the Donor and FA Labeled Human IL-6 R alpha as the Acceptor.
— When the sample does not contain the inhibitors that block the binding of human IL-6 to human IL-6 R alpha, the Donor and Acceptor are in close proximity because of the binding of human IL-6 and human IL-6 R alpha. Upon Donor excitation with light of a specific wavelength (337nm), in addition to Donor emission (620nm), non-radiative transfer of energy occurs between Donor and Acceptor, resulting in Acceptor emission (665nm).
— When the sample contains the inhibitors that block the binding of human IL-6 to human IL-6 R alpha, the components inhibit the binding between the Donor and Acceptor and thereby prevent FRET from occurring.
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Performance Data
Bioactivity-TR-FRET
Please refer to DS document for the assay protocol.
In this TR-FRET assay, Human IL-6 Protein Europium-chelate is used as the Donor and FA Labeled Human IL-6 R alpha is used as the Acceptor. An inhibition Assay was performed to evaluate the interaction between human IL-6 and human IL-6 R alpha using Anti- IL-6 Neutralizing Antibody resulting in a typical IC50 of 0.4213 nM (QC tested).
The kit is suitable for the detection and characterization of IL-6 / IL-6 R alpha inhibitors. It was shown that the two anti-IL-6 antibodies (FRTP023-C03 & Siltuximab) disrupted the interaction, with an IC50 of 0.4261 & 0.4943 nM. The anti-IL-6 R antibody (Tocilizumab) disrupted the interaction, with an IC50 of 3.348 nM. The anti-TNF-alpha (Adalimumab) showed no significant inhibitory effect in the assay as expected.
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