Protein stains for proteomic applications: which, when, why?
This review recollects literature data on sensitivity and dynamic range for the most commonly used colorimetric and fluorescent dyes for general protein staining, and summarizes procedures for the most common PTM-specific detection methods. It also compiles some important points to be considered in imaging and evaluation. In addition to theoretical considerations, examples are provided to illustrate differential staining of specific proteins with different detection methods. This includes a large body of original data on the comparative evaluation of several pre- and post-electrophoresis stains used in parallel on a single specimen, horse serum run in 2-DE (IPG-DALT). A number of proteins/protein spots are found to be over- or under-revealed with some of the staining procedures.
Miller
Ingrid
I
Institut für Medizinische Chemie, Department für Naturwissenschaften, Veterinärmedizinische Universität Wien, Wien, Austria. ingrid.miller@vu-wien.ac.at
Crawford
Johanne
J
Gianazza
Elisabetta
E
D003124:Colorimetry / Q000295:instrumentation / Q000379:methods; D003545:Cysteine / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D005456:Fluorescent Dyes / Q000494:pharmacology; D006023:Glycoproteins / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007091:Image Processing, Computer-Assisted; D008239:Lysine / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D040901:Proteomics / Q000295:instrumentation* / Q000379:methods*; D012394:Rosaniline Dyes / Q000494:pharmacology; D012680:Sensitivity and Specificity; D012835:Silver Nitrate / Q000494:pharmacology; D013053:Spectrophotometry
D005456:Fluorescent Dyes; D006023:Glycoproteins; D011506:Proteins; D012394:Rosaniline Dyes; C048139:Coomassie blue; D012835:Silver Nitrate; C035906:Fast Green; D008239:Lysine; D003545:Cysteine