Promega, VB4050, Lumit® Immunoassay Labeling Kit and Detection Reagents

How the Lumit® Immunoassay Labeling Kit and Detection Reagents Work
Lumit® Immunoassays are based on the chemical labeling of antibodies and proteins with SmBiT and LgBiT using HaloTag® technology. In the presence of an analyte, the two antibodies come into close proximity and allow SmBiT and LgBiT to form an active enzyme and generate a bright luminescence signal proportional to analyte concentration.
Lumit® Detection Reagents A and B are specially designed to detect luminescence signal from Lumit® Immunoassays when using labeled primary antibodies. Lumit® Detection Reagent A is used with biochemical and cell lysates, while Lumit® Detection Reagent B is specifically designed for analyte detection in cell culture media containing Fetal Bovine Serum (FBS) and for analyte detection directly in cell culture wells containing media and cells.
Chemical Labeling Using HaloTag® Technology
Labeling is a two-step process, where amine-reactive HaloTag® Succinimidyl Ester (O4) Ligand reacts with primary amines on lysine amino acids on the antibodies and proteins. Antibodies and proteins labeled with the HaloTag® ligand are then incubated with HaloTag®-LgBiT or HaloTag®-SmBiT fusion protein to make a covalent conjugate of antibody-HaloTag®-LgBiT or antibody-HaloTag®-SmBiT.
Simple Add-and-Mix Protocol, No Wash Steps Required!
The Lumit® Immunoassay does not require media removal or lysate transfer. The protocol simply involves addition of the sample to the labeled antibodies, addition of the detection reagent and reading of the luminescent signal—all in a single plate!