摘要
目的对人乳牙牙髓干细胞全蛋白质进行非胶分级分离电泳,并用液质联用质谱分析所得蛋白质组分。方法对人乳牙牙髓干细胞进行裂解,将所得的细胞全蛋白质均分到12孔样品槽里进行非胶分级分离电泳,液相回收电泳后的12个组分,对各组分进行SDS-PAGE分离,并对各组分进行溶液酶切后再进行液质联用质谱分析。结果 SDS-PAGE图谱显示人乳牙牙髓干细胞全蛋白质经非胶分级分离电泳分成了12个不同的蛋白质组分,液质联用对各组分进行分析均成功鉴定到一定数目蛋白质。结论非胶分级分离电泳能将复杂蛋白质样品进行预分离,液相回收分离后样品便于进行液质联用分析。
Objective To isolate and identify the total cell proteins of stem cells from human exfoliated deciduous teeth by OFFGEL electrophoresis and liquid chromatography with mass spectrometry( MS). Methods The total cell proteins were extracted from stem cells from human exfoliated deciduous teeth,and assigned into 12 sample wells for OFFGEL electrophoresis( OGE). The 12 protein fractions were obtained by liquid recovery,then isolated by SDS-PAGE and digested to the peptides,and finally identified by nano liquid chromatography-electrospray ionization-MS / MS( LC-ESI-MS / MS). Results The map of SDS-PAGE revealed that the total cell proteins of stem cells from human exfoliated deciduous teeth were composed of 12 different fractions after OGE. A number of proteins from the12 fractions were successfully identified by nano LC-ESI-MS / MS. Conclusion The complex protein samples can be isolated,and the separated proteins are easy to be identified by LC-MS / MS.
出处
《山西医科大学学报》
CAS
2016年第3期250-254,共5页
Journal of Shanxi Medical University
基金
首都医科大学科研基金资助项目(校技术类2014JS04)