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A novel [Ag(NH 3 ) 2 ] + probe for chemiluminescent imaging detection of proteins after polyacrylamide gel electrophoresis
Author(s) -
Xiong Xin,
Wang Zhenzhen,
Baeyens Willy R. G.,
Delanghe Joris R.,
Huang Zhi,
Huang Guangming,
Ouyang Jin
Publication year - 2007
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200700092
Subject(s) - chemistry , reagent , chemiluminescence , chromatography , polyacrylamide , detection limit , polyacrylamide gel electrophoresis , luminol , silver stain , staining , ammonium persulfate , gel electrophoresis , chromogenic , electrophoresis , biochemistry , microbiology and biotechnology , biology , enzyme , genetics , polymer chemistry , organic chemistry , polymerization , polymer
The development of a novel [Ag(NH 3 ) 2 ] + probe chemiluminescence (CL)‐based imaging method for the detection of various proteins after PAGE is described. The detection is based upon the probe [Ag(NH 3 ) 2 ] + catalyzing the CL reaction of the luminol–potassium persulfate system. The proposed method detects various proteins labeled by [Ag(NH 3 ) 2 ] + and expands the application scope to SDS gels. It also detects proteins directly in polyacrylamide gels, without tedious transferring procedures. Furthermore, successful identification of proteins by peptide mass profiling using ionization MS was easily performed, and no pretreatments of gel prior to digestion are needed. Detection limits for standard marker proteins match CBB‐R250 staining and the linear dynamic range is superior to CBB‐R250 staining and silver staining. The CL imaging conditions, including luminescent reagents, silver ion concentration, the ammonia‐controlled system and the washing reagents parameters have also been optimized.