Preparation of protein extracts from yeast.
Three protocols are presented for preparing protein extracts; they differ primarily in the way the cells are broken. In the basic protocol, cells are enzymatically converted to spheroplasts, which are then lysed by a combination of osmotic shock and Dounce homogenization. A support protocol for isolating intact nuclei by differential centrifugation is also presented. An alternate protocol describes mechanical breakage of cells by vortexing in the presence of glass beads. In a second alternate protocol, growing cells are frozen immediately in liquid nitrogen and then lysed by grinding in an industrial-strength blender in the presence of liquid nitrogen.
Dunn
B
B
Memorial Sloan-Kettering Cancer Center, New York, New York, USA.
Wobbe
C R
CR
D002467:Cell Nucleus / Q000737:chemistry; D002498:Centrifugation; D005656:Fungal Proteins / Q000302:isolation & purification*; D005898:Glass; D009584:Nitrogen; D013104:Spheroplasts / Q000302:isolation & purification*; D015003:Yeasts
D005656:Fungal Proteins; D009584:Nitrogen