Cite as: Cold Spring Harb. Protoc.; 2009; doi:10.1101/pdb.prot5110

This Protocol
Right arrow Full Text
Right arrow Update/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 Similar articles in PubMed
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 Google Scholar
Google Scholar
Right arrow Articles by Link, A. J.
Right arrow Articles by LaBaer, J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Link, A. J.
Right arrow Articles by LaBaer, J.
Related Collections
Right arrow CSHL Proteomics Course
Right arrow Protein Identification and Analysis
Right arrow Proteins and Proteomics, general
Right arrow Mass Spectrometry
Right arrow Protein Classification and Structure Prediction
Right arrow Electrophoresis, general
Right arrow Electrophoresis of Proteins
Right arrowRelated Protocols
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

In-Gel Trypsin Digest of Gel-Fractionated Proteins

Andrew J. Link1,3 and Joshua LaBaer2

1Vanderbilt University School of Medicine, Nashville, TN 37232-8575, USA
2Harvard University School of Medicine, Harvard Institute of Proteomics, Cambridge, MA 02141-2023, USA

3Corresponding author (andrew.link{at}vanderbilt.edu)


INTRODUCTION

This protocol describes a method for in-gel digestion of proteins after fractionation using SDS-PAGE. It is applicable to both one- and two-dimensional polyacrylamide gels of different thicknesses, acrylamide concentrations, and band (spot) sizes. This protocol followed by liquid chromatography/mass spectrometry/mass spectrometry (LC-MS/MS) or matrix-assisted laser desorption/ionization/time of flight/time of flight (MALDI-TOF/TOF) is capable of identifying low femtomole quantities of a protein. High-sensitivity mass spectrometry should be capable of identifying attomole quantities of protein.


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?

Related Protocols

SDS-PAGE of Proteins
Richard J. Simpson
Cold Spring Harb Protoc 2006: 4313. [Extract] [Full Text]

SDS-Polyacrylamide Gel Electrophoresis of Proteins
Joseph Sambrook and David W. Russell
Cold Spring Harb Protoc 2006: 4540. [Abstract] [Full Text]