Check out the video below showing how to mix LCP:
Lipidic cubic phase is usually made up with a protein to monoolein ratio of 2:3 (40%:60%).
LCP mixing adapter (MD6-17), pipette & tips, two gastight Hamilton syringes (250 µL), plungers, needles (big inner and big outer diameter 22 gauge should be fine), small/large connector, two plastic screws and tube, monoolein (melted), protein solution.
- Fill one of the Hamilton syringes with the protein solution (eg. 40 µL) using a needle. Attach the large connector to the syringe, then insert the syringe into the large connector tube and screw tightly.
- Melt the monoolein at ~40-50°C.
- The other syringe is filled with monoolein in the following way: cut off the end of the pipette tip (to make it easier to work with the viscous monoolein) and draw up 60% (e.g. 60 µL) of melted monoolein. The monoolein is then pipetted into the syringe whilst simultaneously pulling the syringe plunger (see movie for more detailed instructions). This has to be done rather quickly since the monoolein otherwise will become stiff. Then insert the syringe into the other end of the connector tube and seal it tight with the second screw.
- Mixing: protein solution and monoolein are mixed through moving the plungers from side to side until the phase is clear (not cloudy) and homogenous (this will be the cubic phase).
- The cubic phase can then be transferred into a suitable container (eg. glass tube) for crystal trials.
A screen specially designed to to work in synergy with Lipidic Cubic Phase should be used. Developed in conjunction with the successful laboratory of Prof. Osamu Nureki at the University of Tokyo, Japan, MemMeso™ has been used to solve numerous crystal structures of membrane proteins, including Channelrhodopsin. See the full range here.
We also suggest the Wizard Cubic™ LCP screen was designed specifically for the LCP method, and comes in HT-96 format.
Our Laminex™ sandwich plates provide great optical quality, and are made with two different spacer sizes. The most popular size is the 100 micron thickness so we suggest you start with this. The starter kit comes with a full range of sealing covers for you to try out. See the full range of LCP plates here.
If using an automated imaging system such as the Rock Imager, we also stock HT versions.
Not sure which plates to choose? Have a look at this article
To harvest your crystals, we provide a tungsten carbide scriber for opening up the glass sandwich plates, and fine tweezers - more information here...
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