LCP Tools >> LCP-FRAP Station
LCP-FRAP Station
An LCP-FRAP assay was developed to rapidly screen diffusion properties of membrane proteins in LCP under variety of crystallization conditions (Cherezov et al., 2008; Xu et al., 2011). The assay is being used to pre-screen multiple protein constructs, lipids and additives to eliminate those conditions that are non-conducive to protein diffusion, and thus crystallization, from the subsequent crystallization trials. An automated LCP-FRAP station consists of the following components:
- Zeiss AxioImager A1 microscope with HBO 100 illumination and 5x, 10x, 40x and 100x objectives.
- Fluorescence filters for Bimane, Fluorescein, Rhodamine 6G and Cy3 dyes.
- Micropoint dye cell laser with tunable wavelength (Photonic Instruments).
- Cooled (-30 °C) CCD (1392 x 1040 pixels, 6.45 µm/pixel) 14 bit monochrome FireWire camera CoolSnap HQ2 (Photometrics).
- MS-2000 XYZ microscope stage with an SBS-type plate adapter and controller (ASI).
- FilterWheel and Shutter with controller.
- Image-Pro Advanced Microscopy suit (Media Cybernetics) to control and automate data acquisition.
A commercial version of the LCP-FRAP station is available from Formulatrix.
REFERENCES:
Cherezov, V., Liu, J., Griffith, M., Hanson, M.A. and R. C. Stevens (2008) LCP-FRAP Assay for Pre-Screening Membrane Proteins for in Meso Crystallization. Cryst Growth Des 8: 4307-4315. >>
Xu, F., Liu, W., Hanson, M.A., Stevens, R.C. and V. Cherezov (2011) Development of an automated high throughput LCP-FRAP assay to guide membrane protein crystallization in lipid mesophases. Cryst Growth Des 11: 1193-1201. >>