Human Fc gamma RIIIA / CD16a (V176) binding Kit (TR-FRET)

Human Fc gamma RIIIA / CD16a (V176) binding Kit (TR-FRET)

• Background: CD16 is a low affinity Fc receptor, and has been identified as Fc receptors FcγRIIIa (CD16a) and FcγRIIIb (CD16b). These receptors bind to the Fc portion of IgG antibodies. CD16 encoded by two different highly homologous genes in a cell type-specific manner.CD16 is found on the surface of natural killer cells, neutrophil polymorphonuclear leukocytes, monocytes and macrophages. CD16a antigen is also known as Low affinity immunoglobulin gamma Fc region receptor III-A, Fc-gamma RIII-alpha. CD16b is a low-affinity, GPI-linked receptor expressed by neutrophils and eosinophils, whereas CD16a is an intermediate affinity polypeptide-anchored transmembrane glycoprotein expressed natural killer cells, macrophages, subpopulation of T-cells, immature thymocytes and placentaltrophoblasts.CD16a is involved in phagocytosis, secretion of enzymes and inflammatory mediators, antibody­dependent cytotoxicity and clearance of immune complexes. Aberrant expression or mutations of CD16a is implicated in susceptibility to recurrent viral infections, systemic lupus erythematosus, and alloimmune neonatal neutropenia.
• Description: CD16 is a low affinity Fc receptor, and has been identified as Fc receptors FcγRIIIa (CD16a) and FcγRIIIb (CD16b). These receptors bind to the Fc portion of IgG antibodies. CD16 encoded by two different highly homologous genes in a cell type-specific manner.CD16 is found on the surface of natural killer cells, neutrophil polymorphonuclear leukocytes, monocytes and macrophages. CD16a antigen is also known as Low affinity immunoglobulin gamma Fc region receptor III-A, Fc-gamma RIII-alpha. CD16b is a low-affinity, GPI-linked receptor expressed by neutrophils and eosinophils, whereas CD16a is an intermediate affinity polypeptide-anchored transmembrane glycoprotein expressed natural killer cells, macrophages, subpopulation of T-cells, immature thymocytes and placentaltrophoblasts.CD16a is involved in phagocytosis, secretion of enzymes and inflammatory mediators, antibody­dependent cytotoxicity and clearance of immune complexes. Aberrant expression or mutations of CD16a is implicated in susceptibility to recurrent viral infections, systemic lupus erythematosus, and alloimmune neonatal neutropenia.
• Reactivity: Human
• Target: Fc gamma RIIIA / CD16a
• Applications: The kit is developed for the binding of IgG Fc region to the human Fc gamma RIIIA / CD16a (V176) receptor. It is used as a universal detection tool to identifying the ability of antibody drugs to bind to Fc gamma RIIIA / CD16a (V176).
• 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 RIIIA / CD16a (V176) binding kit (TR-FRET) is based on TR-FRET technology (Time-Resolved Fluorescence Resonance Energy Transfer). Use the mixture of biotinylated human Fc gamma RIIIA / CD16a (V176) 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 RIIIA / CD16a (V176) 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 = "Signal 665 nm" / "Signal 620 nm" × 10^4. The Ratio value is negatively correlated with the antibody content in the sample. - When the sample does not contain human Fc gamma RIIIA / CD16a (V176) binding components, the donor and acceptor are in close proximity because of the binding of human Fc gamma RIIIA / CD16a (V176) 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 RIIIA / CD16a (V176) 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