DMPC:CHAPSO Pre-Mixed Bicelles

Crystallization of membrane proteins is a challenging undertaking and a number of alternative techniques have been developed such as lipidic cubic phase and bicelle crystallization. Both of these techniques allow for the reconstitution of a detergent solubilized membrane protein in a lipidic environment, and the proteins tend to pack into type-I crystals. Bicelles are formed from a mixture of 1,2-dimyristoyl- sn -glycero-3-phosphocholine (DMPC, D514) and 1,2-dihexanoyl- sn -glycero-3-phosphocholine (DHPC, D606) or a mixture of DMPC and 3-((3-cholamidopropyl)dimethylammonio)-2-hydroxy-1-propanesulfonate (CHAPSO, C317). Bicelles are bilayer discs with the bilayer formed by the lipid (DMPC) and the detergent (DHPC or CHAPSO) covering the edges of the hydrophobic bilayer.
 
Setting up a crystallization trial in bicelles can be done with four simple steps:

• Preparation of the bicelle medium
• Incorporation of the protein into bicelles
• Setup of crystallization trials
• Monitoring, harvesting, and data collection of crystals

To simplify this process, Anatrace offers premixed bicelle solutions. These solutions contain 0.25 ml of a 30% solution of either DMPC:CHAPSO or DMPC:DHPC bicelles at a 2.8:1 ratio. For initial crystallization screening, we recommend screening between 5% - 8% bicelles. To reconstitute a protein into bicelles, protein is mixed with the bicelle solution in a 4:1 ratio. For example, to create 100 µl of a 5% protein-bicelle mixture, the 30% stock bicelle solution is first diluted with H₂O to 25% (16.7 µl 30% bicelles + 3.3 µl H₂O = 20 µl 25% bicelles), and then added to 80 µl of concentrated protein solution. The reconstituted protein in bicelles can then be used to setup crystallization experiments in either hanging drop, sitting drop, or the Microlytic Crystal Former plates. A full protocol for protein reconstitution and crystallization is be available under the Technical Documentation tab.

Note: Bicelles are shipped at room temperature. Store at -20^oC upon receiving. 

Citations:
1.) Faham, S. and Bowie, J. U. (2002) J. Mol. Biol. 316(1), 1-6.
2.) Poulos, S. et al. (2015) Methods in Enzymology 557, 393-416.
3.) Kimble-Hill, A. C. (2013) Front. Biol. 8(3), 261-272.
4.) Agah, S. and Faham, S. (2012) Methods in Molecular Biology 914, 3-16.

• Bicelle Crystallization Protocol (PDF)

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