Cite as: Cold Spring Harb. Protoc.; 2010; doi:10.1101/pdb.prot5379

This Protocol
Right arrow Full Text
Right arrow Discuss this protocolDiscussion icon
Right arrow Alert me when this protocol is cited
Right arrow Alert me when comments are published
Right arrow Alert me if a correction is posted
Services
Right arrow Similar protocols in this database
Right arrow Alert me to new releases of protocols
Right arrow Save to Personal Folders
Right arrow Download to citation manager
Right arrow Printer-friendly versionPrinter-friendly version
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Mohammad, J.
Right arrow Search for Related Content
PubMed
Right arrow Articles by Mohammad, J.
Related Collections
Right arrow Protein Identification and Analysis
Right arrow Proteins and Proteomics, general
Right arrow Chromatography
Right arrow Chromatography, general
Right arrow Chromatofocusing
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

protocolProtocol

Cleaning, Regeneration, and Storage of Chromatofocusing Columns

Jamil Mohammad

Adapted from Purifying Proteins for Proteomics (ed. Simpson). CSHL Press, Cold Spring Harbor, NY, USA, 2004.


INTRODUCTION

Chromatofocusing separates proteins on the basis of differences in their isoelectric point (pI). A chromatofocusing column should be cleaned and regenerated after use, as described in this protocol, to remove any strongly bound substances. The following observations indicate that column cleaning may be necessary: increased back pressure in the column, a color change at the top of the column, loss of resolution, or reduced sample recovery. To avoid these problems, clean the column routinely (e.g., after every fifth or tenth run). More frequent and extended cleaning procedures may be necessary if complex samples are applied. The steps in this article are guidelines. Personal experience with possible contamination should be used to evaluate which cleaning method is suitable.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?


This article has been cited by other articles:


Home page
 Cold Spring Harb ProtocHome page
J. Mohammad
Chromatofocusing
Cold Spring Harb Protoc, February 1, 2010; 2010(2): 10.1101/pdb.top67.
[Abstract] [Full Text]

Home page
 Cold Spring Harb ProtocHome page
J. Mohammad
Preparation of the Gel and Column for Chromatofocusing
Cold Spring Harb Protoc, February 1, 2010; 2010(2): 10.1101/pdb.prot5375.
[Abstract] [Full Text]

Home page
 Cold Spring Harb ProtocHome page
J. Mohammad
Preparation of the Sample for Chromatofocusing: Buffer Exchange and Desalting
Cold Spring Harb Protoc, February 1, 2010; 2010(2): 10.1101/pdb.prot5376.
[Abstract] [Full Text]

Home page
 Cold Spring Harb ProtocHome page
J. Mohammad
Purification of Proteins by Chromatofocusing
Cold Spring Harb Protoc, February 1, 2010; 2010(2): 10.1101/pdb.prot5377.
[Abstract] [Full Text]

Home page
 Cold Spring Harb ProtocHome page
J. Mohammad
Removal of Elution Buffer from Protein Sample Fractions after Chromatofocusing
Cold Spring Harb Protoc, February 1, 2010; 2010(2): 10.1101/pdb.prot5378.
[Abstract] [Full Text]