膜杰作
Star Staining
优势与特点（FAB）
- Cost effective - Sufficient quantity at a lower price, accounting for dilution and pipetting losses.
- Comprehensive validation - Validated with various antibody subtypes and 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 - Supports 500 tests, ideal for high-throughput screening.
- Fast completion - Results in just 1 hour.
产品参数（Product Specifications）
| Assay Type | Inhibition-TR-FRET |
| Analyte | Human IgG, Human IgG Fc protein, Anti-human CD32a antibody |
| Format | 100T/500T |
| Reactivity | Human |
| Regulatory Status | RUO |
| Sensitivity | IC50=816.6nM |
| Standard Curve Range | 2.4414 nM-10000 nM |
| Assay Time | 1 hr |
| Suitable Sample Type | For the binding of IgG Fc region to the human CD32a |
| Sample volume | 10 μL |
产品概述（Product Overview）
The Human Fc gamma RIIA / CD32a (R167) 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 CD32a (R167) and antibody drug candidates or CD32a (R167) inhibitors. It is designed to facilitate the ADCC and ADCP functional performance evaluation of antibody drug candidates, high-throughput screening of CD32a (R167) 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 CD32a (R167).
存储（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.
组分（Materials Provided）
| ID | Components | Size |
| FRT05-C01 | Human Fc gamma RIIA / CD32a (R167) Protein Europium-chelate | 100 tests/500 tests |
| FRT05-C02 | FA labeled human IgG antibody | 100 tests/500 tests |
| FRT05-C03 | Human IgG Standard | 400 μg/100 tests
2000 μg/500 tests |
| FRT05-C04 | Sample Dilution Buffer | 10 mL/100tests & 500tests |
| FRT05-C05 | Detection Buffer | 10 mL/100tests & 500tests |
原理（Assay Principles）
Human Fc gamma RIIA / CD32a (R167) binding kit (TR-FRET) is based on TR-FRET technology (Time-Resolved Fluorescence Resonance Energy Transfer). Use the mixture of biotinylated human Fc gamma RIIA / CD32a (R167) and Europium-chelate labeled streptavidin as the donor, FA labeled Human IgG1 antibody as the acceptor.
- In the absence of human Fc gamma RIIA/CD32a (R167) binding components, the donor and acceptor are in close proximity due to the binding of human Fc gamma RIIA/CD32a (R167) 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 RIIA/CD32a (R167) binding components, they disrupt the donor-acceptor interaction, preventing FRET from occurring.
质量管理控制体系（QMS）
活性（Bioactivity）-TR-FRET Please refer to DS document for the assay protocol.
Inhibition Assay of interaction of Europium-chelate labeled human Fc gamma RIIA / CD32a (R167) and FA labeled human lgG by Human IgG standard in a homogeneous (no wash) TR-FRET (Time-Resolved Fluorescence Resonance Energy Transfer) competition assay, with a typical IC50 of 913 nM (QC tested).
The kit has been used to detect different subclasses of Human IgG (Human IgG1, Human IgG2, Human IgG3 and Human IgG4), which exhibit different IC50 results as expected. As shown in the following figure, human CD32a (R167) binds to human IgG1, IgG2, IgG3 and IgG4 with low affinity.
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 CD32a (R167) binds to mouse IgG1, mouse IgG2a, and mouse IgG2b with low affinity as observed, and mouse IgG1 show the higher affinity than mouse IgG2a or mouse IgG2b.
The kit has been used to detect four FDA approved antibody drugs with different affinities binding to human CD32a (R167). Bevacizumab, Toripalimab, Eculizumab and Efgartigimod alfa bind to human CD32a (R167) with the low affinity from 1 μM to 10 μM.
Verify potential matrix effects by adding different levels of DEME, RPMI1640, FBS and HSA to the Sample Diluted buffer.
产品推荐（Recommended Products）
背景（Background）：Fc gamma RIIA / CD32a
Receptors for the Fc region of IgG (Fc γ R) are members of the Ig superfamily that function in the activation or inhibition of immune responses. Three classes of human Fc γ Rs: RI (CD64), RII (CD32), and RIII (CD16), which generate multiple isoforms, are recognized.
There are three genes for human Fcγ RII /CD32 (A, B, and C) and one for mouse Fcγ RII B (CD32B). CD32 is a low affinity receptor for IgG. The activating isoform, CD32A, is expressed on monocytes, neutrophils, platelets and dendritic cells. CD32A is expressed on many immune cell types (macrophage, neutrophil, eosinophils, platelets, dendritic cells and Langerhan cells), where inhibitory ITIMbearing receptors may also be coexpressed and coengaged by specific ligands. CD32A delivers an activating signal upon ligand binding, and results in the initiation of inflammatory responses including cytolysis, phagocytosis, degranulation and cytokine production. The responses can be modulated by signals from the coexpressed inhibitory receptors such as CD32B
