Human Fc gamma RIIA / CD32a (H167) binding Kit (TR-FRET)

Human Fc gamma RIIA / CD32a (H167) binding Kit (TR-FRET)

• Background: 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, and the strength of the signal is dependent on the ratio of expression of the activating and inhibitory receptors.
• Description: 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, and the strength of the signal is dependent on the ratio of expression of the activating and inhibitory receptors.
• Reactivity: Human
• Target: Fc gamma RIIA / CD32a
• Applications: The kit is developed for the binding of IgG Fc region to the human Fc gamma RIIA / CD32a (H167) receptor. It is used as a universal detection tool to identifying the ability of antibody drugs to bind to Fc gamma RIIA / CD32a (H167).
• Stability: 1. Unopened kit should be stored at 2°C-8°C 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.
• Format: Solid
• Components: Human Fc gamma RIIA / CD32a (H167) 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 (H167) and Europium-chelate labeled streptavidin as the donor, FA labeled Human IgG1 antibody as the acceptor. Your experiment will include 3 simple steps: 1)Mix the sample or Human IgG standard in the kit with Human Fc gamma RIIA / CD32a (H167) Protein Europium-chelate (Donor) and incubate at room temperature for 0.5 hours. 2)Add FA labeled human IgG antibody (Acceptor) and incubate at room temperature for at least 0.5 hours. 3)Use the TR-FRET module of a microplate reader to read the fluorescence signal at 665nm and 620nm. Calculate the Ratio based on the formula Ratio = × . The Ratio value is negatively correlated with the antibody content in the sample. - When the sample does not contain human Fc gamma RIIA / CD32a (H167) binding components, the donor and acceptor are in close proximity because of the binding of human Fc gamma RIIA / CD32a (H167) and FA labeled Human IgG1 antibody. The 620nm signal emitted by the donor under specific light source excitation is received by the acceptor, emitting a 665nm signal. - When the sample contains human Fc gamma RIIA / CD32a (H167) binding components, the components inhibit the binding between the donor and acceptor and thereby prevents FRET from occurring.
• Shipping Conditions: Blue ice
• Storage Conditions: 2-8°C