How does biotin label transfer work?
The label-transfer method involves transfer of a biotin molecule from a purified bait protein to a binding partner designated as the prey protein. Using these reagents, a -SH-containing bait protein is first biotinylated via the Mts moiety. After removal of excess reagent and incubation in the dark with a putative prey protein or prey protein-containing lysate, the Atf moiety is activated with UV light capturing the interacting complex. Reduction of the disulfide bond formed by reaction of -SH with the Mts group of the label-transfer reagent results in the transfer of the biotin label from the bait protein to the prey protein. The biotin can be used to both enrich for and detect the interacting complex or prey protein using immobilized Monomeric Avidin or Streptavidin-HRP-based detection, respectively (Figure 1). Figure 1. Label-transfer scheme. The bait protein is first biotinylated through the Mts moiety. An interacting prey protein is then captured by photoactivation of the Atf moie
