Primary antibodies to the target are used to detect the cytokine. The antibodies, selected for their specific and sensitive detection of the targets, are labeled with the LgBiT and SmBiT subunits of NanoBiT® Luciferase. In the presence of the target, the subunits are brought together to reconstitute active luciferase enzyme. Upon addition of the optimized substrate, a bright luminescent signal is generated.
The protocol simply requires addition of the labeled antibodies to the sample, addition of the detection reagent and reading of the luminescent signal. The entire protocol is completed in less than 70 minutes.