[Identification of phosphoproteome in mice neurons by isotope-labeling technique combining with chemiluminescence Western blotting method].
This study was aimed to work out a simple, applicable, sensitive and specific protocol for the identification of phosphoproteome. Isotope-labeling, two-dimensional electrophoresis, autoradiography and so on were used to establish a phosphoproteome map of mice neurons, and then chemiluminescence Western blotting was utilized to detect three phosphoproteins PI3Kr3, MEK1 and PKCalpha selectively. The results of comparison showed that the blots of PI3Kr3, MEK1 and PKCalpha on autoradiography map were almost identical with the blots of PI3Kr3, MEK1 and PKCalpha on chemiluminescence Western blotting maps. So this protocol based on isotope labeling and chemiluminescence Western blotting methods has proven to be sensitive and specific in the identification of phosphoproteome.
Yao
Fuli
F
Department of Biochemistry, School of Basic Medical Sciences, Luzhou Medical College, Luzhou 646000, China.
Li
Hong
H
Yin
Kang
K
He
Tao
T
Wu
Minglu
M
Xiao
Bin
B
D000818:Animals; D000831:Animals, Newborn; D015153:Blotting, Western / Q000379:methods; D001921:Brain / Q000166:cytology; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D008163:Luminescent Measurements / Q000379:methods; D051379:Mice; D009474:Neurons / Q000737:chemistry* / Q000166:cytology; D010750:Phosphoproteins / Q000032:analysis*; D010761:Phosphorus Radioisotopes; D020543:Proteome / Q000032:analysis*; D012680:Sensitivity and Specificity
D010750:Phosphoproteins; D010761:Phosphorus Radioisotopes; D020543:Proteome