Abstract
Western blotting is a well established analytical technique used to detect specific proteins in a sample1–10. Following protein separation through gel electrophoresis, the proteins are transferred to a solid membrane support. Once on the membrane, a protein of interest can be detected with antibodies specific for the target protein5,6. Numerous molecular biology disciplines employ this technology on a daily basis. Although it is a common laboratory practice, western blotting is a time consuming process and technological advances leading to increased time efficiency and quality are invaluable across widespread scientific disciplines.
The Trans-Blot Turbo system reduces the time to perform protein transfers in a high throughput manner, while maintaining high efficiency transfers. The base unit includes an integrated power supply compatible with universal line voltage, as well as an LCD screen and keypad allowing access to operational parameters. The unit has two independently-controlled transfer cassette bays. Each cassette can transfer up to 2 mini gels or a single midi gel simultaneously. The top lid of each transfer cassette is composed of a stainless steel cathode fixed to a spring-loaded plate to ensure that equal and consistent pressure is applied to the transfer sandwich. The bottom tray contains an anode plate made of platinum-coated titanium. From the software interface, optimized preloaded protocols can be selected. Protocols can also be customized and there is space for up to 25 user-defined transfer protocols to be saved and recalled.
While the system can be used with standard blotting reagents for traditional semi-dry transfer, Trans-Blot Turbo transfer packs are necessary for rapid transfer applications. Specialized precut filter paper and PVDF or nitrocellulose membranes are supplied pre-saturated with optimized buffer. The combination of the specialized filter paper and optimized buffer offers higher ion capacity than standard blotting reagents, enabling higher currents to be passed through the transfer stack to speed up protein migration and improve transfer efficiency. Furthermore, there is no need to equilibrate the gel in transfer buffer. When the Trans-Blot Turbo system is used in combination with the Bio-Rad Mini-PROTEAN TGX gels, efficient protein transfer can be performed in three minutes, significantly reducing the time involved in applying transfer technology.
Here, we present a 3-minute transfer protocol, as well as a protocol to simultaneously transfer 4 mini gels in as little as 7 minutes.
