{"product_id":"revvity-64c1qpeg","title":"Revvity, 64C1QPEG, HTRF Human C1q Binding Kit, 500 Assay Points","description":"\u003cp\u003eOverview\u003cbr\u003e\nComplement-Dependent Cytotoxicity (CDC) is an important effector function of antibodies. Activation of the classical complement pathway is initiated by the binding of the complement protein C1q to the Fc region of immunoglobulins bound to a cell surface target antigen. This interaction triggers the sequential enzymatic activation of circulating complement proteins, and finally leads to the formation of the Membrane Attack Complex (MAC) that mediates target cell lysis.\u003cbr\u003e\nThe capacity of C1q binding and resultant CDC activity can influence the safety and efficacy of therapeutic monoclonal antibodies, and therefore requires characterization during the development process. Engineering the Fc region of IgG antibodies (e.g. by mutations or glycoengineering) is currently an active area of investigation to enhance or reduce the ability of biologics to recruit complement.\u003cbr\u003e\nThe human C1q binding kit measures the binding of the Fc region of aggregated IgG antibodies to human C1q in solution.\u003cbr\u003e\nHTRF assays offer many advantages over other technologies:\u003c\/p\u003e\n\n\u003cp\u003eHomogeneous add-and-read format\u003cbr\u003e\nNo wash steps\u003cbr\u003e\nLow background\u003cbr\u003e\nStraightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates\u003cbr\u003e\nStable signal, providing flexibility in time of readout or size of assays\u003cbr\u003e\nHow it works\u003cbr\u003e\nHTRF Human C1q Binding assay principle\u003cbr\u003e\nThe HTRF Human C1q binding assay measures the binding of the Fc region of IgG antibodies to human C1q in buffered solution. Unlike conventional methods that require the immobilization of antibodies on a solid phase, this assay enables simple and rapid characterization of antibodies in a homogeneous generic format. A biotinylated anti-human IgG Fab antibody complexed to streptavidin is used to capture and aggregate the tested antibody in solution. Antibodies bound to human C1q are detected in a sandwich assay format using an anti-C1q antibody labelled with Eu3+ cryptate (donor), and the streptavidin labelled with d2 (acceptor). The interaction between the two partners brings the donor fluorophore into close proximity to the acceptor, thereby generating a FRET signal. Its intensity is directly proportional to the capacity of the antibody Fc region to interact with human C1q.\u003c\/p\u003e\n\n\n\n\n  \u003cbr\u003e\n        \n\n\n\t\u003cbr\u003e\n  \u003cbr\u003e\n        \u003cbr\u003e\n  \n\n      \u003cbr\u003e\n  \u003cbr\u003e\n  \u003cbr\u003e\n    \u003cbr\u003e\n      \u003cbr\u003e\n        \u003cbr\u003e\n        \u003cbr\u003e\n        \u003cbr\u003e\n        \u003cbr\u003e\n      \u003cbr\u003e\n    \n\n    \u003cbr\u003e\n      \n\n      \u003cbr\u003e\n        \n\n      \u003cbr\u003e\n    \n\n  \n\n\n\n\n\u003cp\u003e \u003c\/p\u003e\n\n\n\n\n\u003cp\u003eHTRF Human C1q Binding assay protocol\u003cbr\u003e\nThe human C1q binding assay can be run in a 96- or 384-well low volume white plate (20 µL final). It requires only 1 incubation step after the sequential addition of assay reagents. As described here, tested antibodies (or the standard) are dispensed directly into the assay plate. The premix [Anti-Human IgG-Fab-biotin + Streptavidin-d2] is then added, followed by the dispensing of the human C1q protein and the anti-C1q antibody labelled with Eu3+ Cryptate. After a 3h-incubation at RT, the FRET signal can be recorded on an HTRF compatible reader. No washing steps are needed. The protocol can be further miniaturized or upscaled by simply resizing each addition volume proportionally. Application-Protein-Protein Interaction\u003cbr\u003e\nAutomation Compatible-Yes\u003cbr\u003e\nBrand-HTRF\u003cbr\u003e\nDetection Modality-HTRF\u003cbr\u003e\nProduct Group-Kit\u003cbr\u003e\nSample Volume-5 µL\u003cbr\u003e\nShipping Conditions-Shipped in Dry Ice\u003cbr\u003e\nTarget Class-Binding Assay\u003cbr\u003e\nTarget Species-Human\u003cbr\u003e\nTechnology-TR-FRET\u003cbr\u003e\nTherapeutic Area-Inflammation Oncology \u0026amp; Inflammation\u003cbr\u003e\nUnit Size-500 assay points\u003c\/p\u003e","brand":"Revvity","offers":[{"title":"Default Title","offer_id":46775274012841,"sku":"64C1QPEG","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/revvity-64c1qpeg","provider":"Iright","version":"1.0","type":"link"}