[Feeding mouse with stable isotope labelled with amino acid].
The stable isotope labeling by amino acids in culture (SILAC) based quantitative proteomics serves as a gold standard because of the high accuracy and throughput for protein identifications and quantification. In this study, we discussed the application of SILAC technology in mammal model, and developed quantitative internal standard for comparative proteomics of disease model. The C57BL/6J mice fed by special diet containing the 13C6-Lysine and bred F2 generation. We identified and analyzed total proteins of 9 mice tissues of F2 generation, including brain, lung, heart, stomach, intestine, liver, spleen, kidney, and muscle. Quantitative analysis information could evaluate the mice and different tissues' labeling efficiency. Liver was the most efficient, brain the least, and the labeling efficiency were 96.34%±0.90% and 92.62%±1.98% respectively. The average of the labeling efficiency of F2 generation was 95.80%±0.64%, which met the international standard (≥ 95%) for SILAC quantitative proteomics effective study. SILAC technology was successfully extended to mammalian model system, which will provide powerful tools for the mechanism study of the pathophysiology process with mouse model.
Fengxu
F
Fan
Huiying
H
Gao
Zhongwei
Z
Xu
Linhui
L
Zhai
Tailong
T
Yi
Tao
T
Zhang
Feilin
F
Wu
Chunping
C
Cui
Ping
P
Xu
D000596:Amino Acids / Q000737:chemistry*; D000818:Animals; D004032:Diet / Q000662:veterinary; D007553:Isotope Labeling; D008239:Lysine / Q000737:chemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D011506:Proteins / Q000737:chemistry; D040901:Proteomics / Q000379:methods*
D000596:Amino Acids; D011506:Proteins; D008239:Lysine