Unit

UNIT 10.14 Reversed-Phase Isolation of Peptides

  1. William J. Henzel,
  2. John T. Stults

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb1014s54

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Henzel, W. J. and Stults, J. T. 2001. Reversed-Phase Isolation of Peptides. Current Protocols in Molecular Biology. 54:10.14.1–10.14.17.

Author Information

  1. Genentech, Inc., South San Francisco, California

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: APR 2001

Abstract

In reversed-phase HPLC, peptides are separated on a hydrophobic stationary phase and eluted with a gradient of increasing organic solvent concentration. Protocols describing the separation of peptides in 5- to 500-pmol quantities via narrow-bore (2-mm-i.d.) or microbore (1-mm-i.d.) columns, as well as for the separation of peptides in quantities <5 pmol are provided in this unit. Capillary HPLC columns require a gradient flow rate of 3 to 5 omponents present in a small sample prior to automated sequencing is possible via the procedures for matrix-assisted laser desorption/ionization (MALDI) mass spectrometry and capillary electrophoresis