Preparation and Use of Microcolumns for Sample Desalting or Nanoscale IMAC
This protocol was adapted from “Proteomic Methods for Phosphorylation Site Mapping,” Chapter 9, in Proteins and Proteomics (ed. Simpson). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2003.INTRODUCTION
In this method, microcolumns are prepared using any kind of chromatographic resin. The resin is held in place by making a constriction at the end of a GELoader pipette tip. Sample loading, washing, and elution are performed by loading liquid on top of the resin and applying air pressure to generate a low flow through the column. No frits are necessary.
