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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-5 / IL-5Rα Inhibitor Screening Kit (TR-FRET) is based on a homogeneous (no wash) competition TR-FRET technology (Time-Resolved Fluorescence Resonance Energy Transfer) to screening for inhibitors of human IL-5 binding to human IL-5Rα within 0.5-1 hours. It can also be used as a universal detection tool to identify the ability of human IL-5 to bind to human IL-5Rα.
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
IDComponentsSizeFRTP017-C01Human IL-5 R alpha Protein Europium-chelate100 tests/500 testsFRTP017-C02FA Labeled Human IL-5 Protein100 tests/500 testsFRTP017-C03Monoclonal Anti-IL-5 R alpha 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-5 / IL-5Rα Inhibitor Screening Kit (TR-FRET) is based on TR-FRET technology (Time-Resolved Fluorescence Resonance Energy Transfer). Use a mixture of biotinylated human IL-5 R alpha and Europium-chelate labeled streptavidin as the donor and FA labeled human IL-5 as the acceptor.
— In the absence of inhibitors for human IL-5 R alpha binding to human IL-5, the donor and acceptor are in close proximity due to the binding of human IL-5 R alpha and human IL-5. Upon excitation with a specific light source, the donor emits a 620 nm signal, which is absorbed by the acceptor, resulting in a 665 nm emission.
— In the presence of inhibitors that block the binding of human IL-5 R alpha to human IL-5, the donor-acceptor interaction is disrupted, preventing FRET from occurring.
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Bioactivity-TR-FRET
Please refer to DS document for the assay protocol.
Inhibition of Europium-chelate labeled Human IL-5 R alpha Protein:FA Labeled Human IL-5 binding by Monoclonal Anti-IL-5 R alpha Antibody
Premix serial dilutions of Monoclonal Anti-IL-5 R alpha Antibody (1:2 serial dilution, from 20 μg/mL to 0.02 μg/mL (133.33-0.13 nM)) and Human IL-5 R alpha Protein Europium-chelate. Then add FA Labeled Human IL-5 and incubate at room temperature (20℃-25℃) for 1 hours. Detection was performed with IC50 of 4.323 nM. The assay was performed according to the above-described Datasheet (QC tested).
The kit is suitable for the detection of IL-5 / IL-5Rα inhibitors. It shows that both Benralizumab and Mepolizumab exhibit good inhibitory activity in this TR-FRET competition assay.
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