Navigation auf uzh.ch

Suche

Department of Biochemistry

Lipidic Cubic Phase (LCP)

LCP

The detailed protocol for crystallizing membrane proteins in LCP was published by Martin Caffrey and Vadim Cherezov in: Crystallizing Membrane Proteins Using Lipidic Mesophases, Nat. Protoc. 2009; 4(5): 706–731.

More information about LCP can be found here:
> Vadim Cherezovs USC web site “The Cherezov Lab”

PCC’s standard procedure for preparing the mesophase follows closely the method described by Martin Caffrey and Vadim Cherezov by mixing two parts protein solution (40%) with three parts monoolein (60%; CAS# [111-03-5]; HR2-435). For this procedure two Hamilton syringes (25µL, 50µL or 100µL Syringe Model 1710 RN) get connected by a LCP syringe coupler (Formulatrix, SKU 209526). The mixing process by moving the syringe plungers back and forth is repeated until the cubic mesophase becomes homogenous, clear and does not show any significant birefringence. The protein sample volume required for this procedure is listed in Sample Volume.

The Gryphon-LCP dispenses a 40nl bolus of the LCP-protein mixture and an excess of 800nl precipitant solution in each of the 96 well of a LCP-sandwich plate. The volume of the LCP bolus range between 20nl – 200nl. The SWISSCI LCP sandwich plate used for all LCP-experiments consists of a hydrophobic SBS sized base plate with a 60um or 120um spacer. The experiments are sealed with a glass- or a Laminex Film (MD11-54) cover. PCC recommends for bolus volumes <50nl the sandwich plates with the 60um spacer and the glass cover. The LCP-plate will be incubated at 20°C in the Rock Imager.

Weiterführende Informationen

PCC-Pinboard

New employee at PCC

I am delighted to welcome Görkem Kurtuldu to PCC as a Senior Scientist. She will be stepping into the role as my designated successor. Görkem brings a fresh perspective to PCC, and I am excited to see the positive impact she will make.
Please join me in welcoming Görkem!

New screen:

What’s new:
PCC supports researchers with preparing custom screens through their pipetting service. The researcher can freely compose a screen with up to 30 different stock solutions.
Be sure to have a look and feel free to ask the PCC staff concerning this new service.

We had to adjust the prices for plates that are set up at 4°C due to the significantly higher wear and tear of the liquid handler stationed in the cold room.

The new screen Morpheus® Fusion (Product#: MD1-129) from Molecular Dimensions is now available at PCC as the initial screen GS087.

 

RockImager and Liquid Handler Status

RockImager 20°C:
The imaging schedule is on time.

RockImager 4°C:
The imaging schedule is on time.

Crystal Gryphon 4°C:
In Operation.

Crystal Gryphon LCP 20°C:
In Operation.

Crystal Phoenix 20°C:
In Operation.

Freedom EVO w/ MCA 96 + RoMa
In Operation

Freedom EVO w/ LiHa
In Operation

Contact

Department of Biochemistry
University of Zurich
Winterthurerstr. 190
CH-8057 Zurich
Office: Y44 J 30/34
xtal@bioc.uzh.ch


Görkem Kurtuldu
Tel.: +41 44 635 6557