DELFIA
DELFIA signal generated by addition of enhancement solution.
Dissociation-enhanced lanthanide fluorescence immunoassay (DELFIA) is a common time resolved fluorescent (TRF) high throughput screening method routinely used to assay the phosphorylation of peptide and protein substrates in heterogeneous biochemical kinase assays. In the simplest of these assays, recombinant kinase is added to a biotinylated substrate. The products of the reaction are captured on streptavidin plates, excess reagent is removed by washing the plate, and the phosphorylation of the substrate is assessed using lanthanide-labeled phosphorylation-specific antibodies (direct detection) or unlabeled primary and lanthanide-labeled secondary antibodies (indirect detection). Signal quantification involves the addition of an acidic enhancement solution that dissociates the lanthanide ions from the antibody and allows the formation of stable fluorescent chelates that are quantified by TRF.
Nearly all of Cell Signaling Technology antibodies are available in a formulation without BSA or glycerol for conjugation to lanthanide chelates. Please contact us at drugdiscovery@cellsignal.com for details.
