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Your Position: Home > Kits > Fc gamma RI / CD64 > FRT-03

Human Fc gamma RI / CD64 binding Kit (TR-FRET)

For research use only.

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FAB
  1. Cost effective - Sufficient quantity at a lower price, accounting for dilution and pipetting losses.
  2. Comprehensive validation - Validated with various antibody subtypes and antibody drugs.
  3. Simple and fast operation - No complicated washing steps, significantly reducing time.
  4. High batch consistency - Strict control over raw materials and finished product quality, ensuring a stable supply.
  5. Accurate and reliable results - High sensitivity with minimal matrix effects.
  6. High throughput capability - Supports 500 tests, ideal for high-throughput screening.
  7. Fast completion - Results in just 1 hour.
    Product Specifications
    Assay TypeInhibition-TR-FRET
    AnalyteHuman IgG, Human IgG Fc protein, Anti-human CD64 antibody
    Format100T/500T
    ReactivityHuman
    Regulatory StatusRUO
    SensitivityIC50=5.276nM
    Standard Curve Range0.244 nM-1000 nM
    Assay Time1 hr
    Suitable Sample TypeFor the binding of IgG Fc region to the human CD64
    Sample volume10 μL
  • Product Overview
    Human Fc gamma RI / CD64 binding Kit (TR-FRET) is based on a homogeneous (no wash) competition TR-FRET technology (Time-Resolved Fluorescence Resonance Energy Transfer) to measure the interaction between human CD64 and antibody drug candidates or CD64 inhibitors. It is designed to facilitate the ADCC and ADCP functional performance evaluation of antibody drug candidates, high-throughput screening of CD64 inhibitors within 0.5-1 hours. It can also be used as a universal detection tool to identify the ability of antibody drugs to bind to human CD64.
  • 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
    IDComponentsSize
    FRT03-C01Human Fc gamma RI / CD64 Protein Europium-chelate100 tests/500 tests
    FRT03-C02FA labeled human IgG antibody100 tests/500 tests
    FRT03-C03Human IgG standard100 μg/100 tests
    500 μg/500 tests
    FRT03-C04Sample Dilution Buffer10 mL/100tests & 500tests
    FRT03-C05Detection Buffer10 mL/100tests & 500tests
  • Assay Principles
    This Human Fc gamma RI / CD64 binding kit (TR-FRET) is based on TR-FRET technology (Time-Resolved Fluorescence Resonance Energy Transfer). Use the mixture of biotinylated human Fc gamma RI / CD64 and Europium-chelate labeled streptavidin as the donor, FA labeled Human IgG1 antibody as the acceptor.
    - In the absence of human Fc gamma RI/CD64 binding components, the donor and acceptor are in close proximity due to the binding of human Fc gamma RI/CD64 and FA-labeled Human IgG1 antibody. 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 human Fc gamma RI/CD64 binding components, they disrupt the donor-acceptor interaction, preventing FRET from occurring.

    Assay Principles

Bioactivity-TR-FRET Please refer to DS document for the assay protocol.
 Fc gamma RI / CD64 TR-FRET

Inhibition Assay of interaction of Human CD64 and Human IgG1 antibody by Human IgG standard in a homogeneous (no wash) TR-FRET (Time-Resolved Fluorescence Resonance Energy Transfer) competition assay, with a typical IC50 of 5.276 nM (QC tested).

 Fc gamma RI / CD64 TR-FRET

The kit has been used to detect different subclasses of Human IgG Whole and Fc fragment proteins (Human IgG1, Human IgG2, Human IgG3 and Human IgG4), which exhibit different IC50 results as expected. As shown in the figure, human CD64 is a high affinity receptor that binds to human IgG1, IgG3 and IgG4 with nanomolar affinity, while it has not been shown to bind to human IgG2.

 Fc gamma RI / CD64 TR-FRET

The kit has been used to detect different subclasses of mouse IgG (mouse IgG1, mouse IgG2a and mouse IgG2b), which exhibit different IC50 results as expected. As shown in the figure, human CD64 binds to mouse IgG2a with a high affinity, but not mouse IgG1 and mouse IgG2b.

 Fc gamma RI / CD64 TR-FRET

The kit has been used to detect four FDA approved antibody drugs with different affinities binding to human CD64. Bevacizumab, Toripalimab, and Efgartigimod alfa bind to human CD64 with the nanomolar affinity. The Fc of Eculizumab has been modified into the human IgG2 hinge region and human IgG4 CH2-CH3 region, so it doesn’t bind to human CD64.

 Fc gamma RI / CD64 TR-FRET

Verify potential matrix effects by adding different levels of DEME, RPMI1640, FBS and HSA to the Sample Diluted buffer.

  • Clinical and Translational Updates

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