High throughput pH optimization of protein crystallization.
Most high throughput structural proteomics centers use the sitting-drop method to obtain diffracting crystals for three-dimensional (3D) structure determination of biological macromolecules by x-ray crystallography. Although several robotic systems are available for dispensing the initial sitting-drop screening conditions, generally they are not used for optimization of crystallization conditions. This chapter describes a protocol for such automated systems, which permits easy construction of pH optimization grids using any desired fixed buffer set with varying ionic strengths directly dispensed into the crystallization plate.
Meged
Ran
R
Israel Structural Proteomics Center (ISPC), Department of Structural Biology, Weizmann Institute of Science, Rehovot, Israel.
Dym
Orly
O
Sussman
Joel L
JL
D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods; D006863:Hydrogen-Ion Concentration; D011506:Proteins / Q000737:chemistry*
D011506:Proteins