Products/Services Used | Details | Operation |
---|---|---|
Proteins, Expression, Isolation and Analysis> | … The IVTT solution is prepared through hierarchical addition of 5 μL HeLa cell lysate, 1 μL accessory proteins, 0.8 μL hERG channel-expressing plasmid (Genscript) and 2 μL reaction mix, respectively, to a nuclease-free microtube … | Get A Quote |
The cell membrane is a complex milieu of lipids and proteins. In order to understand the behaviour of individual molecules is it often desirable to examine them as purified components in in vitro systems. Here, we detail the creation and use of droplet interface bilayers (DIBs) which, when coupled to TIRF microscopy, can reveal spatiotemporal and kinetic information for individual membrane proteins. A number of steps are required including modification of the protein sequence to enable the incorporation of appropriate fluorescent labels, expression and purification of the membrane protein and subsequent labelling. Following creation of DIBs, proteins are spontaneously incorporated into the membrane wh... More