Unit

UNIT 13.13 Preparation of Protein Extracts from Yeast

  1. Barbara Dunn1,
  2. C. Richard Wobbe2

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb1313s23

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Dunn, B. and Wobbe, C. R. 2001. Preparation of Protein Extracts from Yeast. Current Protocols in Molecular Biology. 23:IV:13.13:13.13.1–13.13.9.

Author Information

  1. 1

    Memorial Sloan-Kettering Cancer Center, New York, New York

  2. 2

    Harvard Medical School, Boston, Massachusetts

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JUL 1993

Abstract

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.