[Comparison between protein detection methods for two-dimensional gel electrophoresis].
The aim of this study was to compare and analyze protein staining in order to select the optimal staining method for proteomic research. Proteins from acute promyelocytic leukemia cell line NB4 and protein molecular weight marker were separated respectively by 2-D or 1-D electrophoresis and detected respectively by the typical Coomassie brilliant blue, the colloidal Coomassie brilliant blue, the modified Coomassie brilliant blue and the silver staining protocols. The protein detection sensitivity, compatibility with mass spectrometry (MS) and facility of the four staining protocols were compared. The results indicated that the silver staining exhibited the highest sensitivity and MS showed the lowest compatibility 10% of protein identification rate. The detection sensitivity of the modified Coomassie brilliant blue staining was superior to that of other two Coomassie brilliant blue stainings, close to but lower than the silver staining, however the compatibility with MS was better (protein identification rate about 55%). It is concluded that the protein detection sensitivity of the modified Coomassie brilliant blue staining is high, and its compatibility with MS is better, this modified Coomassie brilliant blue staining is an optimal staining method for proteomic research.
Qin
Hui
H
Department of Hematology, State Key Laboratory of Biotherapy for Human Diseases, West China Hospital, Sichuan University, Chengdu 610041, China. qinhui6957@sina100.com
Liu
Ting
T
Liu
Bin
B
Song
Xin
X
Huang
Xin
X
Yang
Jin-Liang
JL
Zhao
Xia
X
Wei
Yue-Quan
YQ
D004396:Coloring Agents; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods; D009363:Neoplasm Proteins / Q000032:analysis*; D012394:Rosaniline Dyes; D016622:Silver Staining; D013194:Staining and Labeling
D004396:Coloring Agents; D009363:Neoplasm Proteins; D012394:Rosaniline Dyes; C004692:coomassie Brilliant Blue