A novel [Ag(NH3)2]+ probe for chemiluminescent imaging detection of proteins after polyacrylamide gel electrophoresis.
The development of a novel [Ag(NH3)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(NH3)2]+ catalyzing the CL reaction of the luminol-potassium persulfate system. The proposed method detects various proteins labeled by [Ag(NH3)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.
Xiong
Xin
X
College of Chemistry, Beijing Normal University, Beijing, PR China.
Wang
Zhenzhen
Z
Baeyens
Willy R G
WR
Delanghe
Joris R
JR
Huang
Zhi
Z
Huang
Guangming
G
Ouyang
Jin
J
D001798:Blood Proteins / Q000032:analysis; D002021:Buffers; D002384:Catalysis; D002712:Chlorides / Q000032:analysis; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D005998:Glycine / Q000737:chemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents / Q000737:chemistry; D007525:Isoelectric Focusing; D007700:Kinetics; D049449:Luminescence; D008163:Luminescent Measurements / Q000379:methods*; D008165:Luminol / Q000737:chemistry; D013058:Mass Spectrometry; D015335:Molecular Probes; D017680:Potassium Compounds / Q000737:chemistry; D011506:Proteins / Q000032:analysis*; D012680:Sensitivity and Specificity; D016622:Silver Staining; D013431:Sulfates / Q000737:chemistry; D013997:Time Factors; D014325:Tromethamine / Q000737:chemistry; D014357:Trypsin / Q000494:pharmacology; D014867:Water / Q000737:chemistry
D001798:Blood Proteins; D002021:Buffers; D002712:Chlorides; D007202:Indicators and Reagents; D015335:Molecular Probes; D017680:Potassium Compounds; D011506:Proteins; D013431:Sulfates; D014325:Tromethamine; D014867:Water; D008165:Luminol; C009007:potassium persulfate; D014357:Trypsin; D005998:Glycine