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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 TNF-alpha / TNFR2 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 TNF-alpha binding to TNFR2 within one hour. It can also be used as a universal detection tool to identify the ability of human TNF-alpha to bind to human TNFR2.
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
IDComponentsSizeFRTP028-C01Human TNF-alpha Protein Europium-chelate100 tests/500 testsFRTP028-C02FA Labeled Human TNFR2 Protein100 tests/500 testsFRTP028-C03Anti-TNF-alpha 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 TNF-alpha / TNFR2 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 TNF-alpha and Europium-chelate labeled streptavidin as the Donor and FA Labeled Human TNFR2 Protein as the Acceptor.
— When the sample does not contain the inhibitors that block the binding of human TNF-alpha to human TNFR2, the Donor and Acceptor are in close proximity because of the binding of human TNF-alpha and human TNFR2. 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 TNF-alpha to human TNFR2, 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 TNF-alpha Protein Europium-chelate is used as the Donor and FA Labeled Human TNFR2 Protein is used as the Acceptor. An inhibition Assay was performed to evaluate the interaction between human TNF-alpha and human TNFR2 using Anti-TNF-alpha Neutralizing Antibody resulting in a typical IC50 of 0.9367 nM (QC tested).
The kit is suitable for the detection and characterization of TNF-alpha/TNFR2 inhibitors. It was shown that the anti-TNF-alpha antibody (Adalimumab) disrupted the interaction, with an IC50 of 0.8714 nM. The anti-TNFR1 antibody (Atrosab) showed no significant inhibitory effect in the assay as expected.
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