| Products/Services Used | Details | Operation |
|---|---|---|
| Bacterial Expression | … coli pcoB gene was synthesized (GenScript), cloned into a pET22 vector with a N-terminal His tag, and transformed into the C43 E. coli strain. 1 L culture in standard LB medium was … | Get A Quote |
Measurement of protein-facilitated copper flux across biological membranes is a considerable challenge. Here, we demonstrate a straightforward microfluidic-derived approach for visualization and measurement of membranous Cu flux. Giant unilamellar vesicles, reconstituted with the membrane protein of interest, are prepared, surface-immobilized, and assessed using a novel quencher-sensor reporter system for detection of copper. With the aid of a syringe pump, the external buffer is exchanged, enabling consistent and precise exchange of solutes, without causing vesicle rupture or uneven local metal concentrations brought about by rapid mixing. This approach bypasses common issues encountered when studying heavy me... More
