Shotgun proteomics: a qualitative approach applying isoelectric focusing on immobilized pH gradient and LC-MS/MS.
Shotgun proteomics is a rapid and near universal strategy to identify proteins in complex protein mixtures. After protein digestion, the resulting peptide mixture is submitted to two orthogonal techniques: peptides are first separated according to their isoelectric point (pI) by isoelectric focusing (IEF) on immobilized pH gradient (IPG); after peptide extraction, they are then separated in the second dimension according to their hydrophobic properties by reverse phase liquid chromatography (RPLC). Finally, they are detected by tandem mass spectrometry (MS/MS) and proteins are matched by means of bioinformatics software.
Swiss Centre for Applied Human Toxicology, Geneva University, Geneva, Switzerland. Laurent.Geiser@unige.ch
Laurent
L
Geiser
Ali R
AR
Vaezzadeh
Jacques M P
JM
Deshusses
Denis F
DF
Hochstrasser
D053000:Analytic Sample Preparation Methods; D002853:Chromatography, Liquid / Q000379:methods*; D056148:Chromatography, Reverse-Phase; D030562:Databases, Protein; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing / Q000379:methods*; D008958:Models, Molecular; D010446:Peptide Fragments / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D011487:Protein Conformation; D011506:Proteins / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
D010446:Peptide Fragments; D011506:Proteins