1-by-1 with HTRF interaction assay is based on Homogeneous Time-Resolved Fluorescence, a highly sensitive and robust Time-Resolved FRET technique. This service allows you to test for direct binding in vitro.
Our offer includes : - Scientific assistance from our protein-interaction experts - Cloning - Protein production and purification: both candidate proteins are produced in bacteria as fusions to different tags - Protein labelling: each protein is labelled either directly or indirectly with anti-tag antibodies coupled to fluorescence donor and acceptor - Competition test: to control that proteins are well recognized by the antibodies and to evaluate the optimal protein concentrations. - HTRF interaction test - Detailed report delivery with protocols, gel pictures, grpahics and comments
Our multi-tag approach and the use of the two donor-acceptor orientations for each protein pair allow for systematically investigating a large panel of combinations, thus increasing the chance of proper protein folding and fluorescence transfer.
Because the assay is homogeneous and requires only low nanomolar concentrations of proteins, multiple interactions can be evaluated simultaneously in a 384-well format.
This makes 1-by-1 with HTRF the perfect output to confirm ULTImate Y2H™ interactions in another system for publication! Our HTRF and Y2H plasmids are fully compatible for direct and efficient subcloning.