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The LANCE® Ultra FCGR3A/CD16a (176Phe/F158) Binding Kit is designed for quantitative determination of Fc-receptor binding using a homogeneous TR-FRET (no-wash steps, no separation steps) assay.
LANCE® and LANCE® (Lanthanide chelate excite) Ultra are our TR-FRET (time-resolved fluorescence resonance energy transfer), homogeneous (no wash) technologies. One protein of interest is labeled with a donor fluorophore (a LANCE Europium chelate) and the second protein is labeled with an acceptor fluorophore [ULight™ dye]. Upon excitation at 320 or 340 nm, energy can be transferred from the donor Europium chelate to the acceptor fluorophore if sufficiently close for FRET (~10 nm). This results in the emission of light at 665 nm.
The Fc-Gamma Receptors (FCGRs) are members of immunoglobulin superfamily and play a critical role in the function of therapeutic antibodies. FCGRs are divided into three classes and FCGR3 (CD16) is expressed as two distinct forms (FCGR3A and FCGR3B) encoded by two different highly homologous genes in a cell type specific manner. FCGR3A is a low/intermediate affinity receptor for polyvalent immune-complexed IgG. It is involved in phagocytosis, secretion of enzymes and inflammatory mediators, antibody-dependent cellular cytotoxicity (ADCC), mast cell degranulation and clearance of immune complexes. In humans, a single nucleotide polymorphism creates two isoforms: high binding (176Val/V158) and low binding (176Phe/F158) forms that, when homozygous, may influence susceptibility to autoimmune diseases or response to therapeutic IgG antibodies. FCGR3A has been considered as an important therapeutic target. This LANCE Ultra assay can be used to determine the binding activity of human IgG Fc fragment to human FCGR3A and also can be used to study how other antibodies bind to FCGR3A by competition assay.
|Assay Target Class||Fc-receptor|
|Detection Method||Time-Resolved Fluorescence (TRF), TR-FRET|
|Experimental Type||In vitro|
|Product Brand Name||LANCE Ultra|
|Shipping Condition||Blue Ice|
|Unit Size||10,000 Assay Points|
This manual explains how to run the LANCE Ultra TR-FRET Fc-receptor binding assay.