摘要
目的:通过对提取狗胰腺组织总蛋白不同方法的研究,建立提取胰腺组织总蛋白的标准方法.方法:用常规的总蛋白一步提取法和三步提取法,以三种溶解性能不同的裂解液分步提取胰腺组织中的总蛋白,测量蛋白浓度并分别用基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption ionization-time of flight-mass spectra,MALDI-TOF-MS)做蛋白全谱.结果:一步法和分步法总蛋白提取量分别为(18.8±1.2)mg和(23.3±0.9)mg,MALDI-TOF-MS蛋白全谱的谱峰数分别为(22.2±4.5)个和(32.8±3.8)个.结论:分步法与常规的一步法相比具有蛋白提取率高、蛋白溶解性好等优点.
AIM： To establish the standard method to extract total protein of pancreatic tissue. METHODS： The three-step se- quential protein extraction method was used to extract the total protein of pancreatic tissue by using three kinds of lysates with different dissolubility, then the protein level was measured and the protein was taken into matrix-assisted laser desorption ionization- time of flight-mass spectra （MALDI-TOF-MS） respectively. RE- SULTS： The extraction amount of total protein with one-step and three-step methods was （ 18.8 ~ 1.2） mg and （23.3 ＋0.9） mg respectively. The number of MALDI-TOF-MS spectra peak was （22.2 ＋ 4.5 ） and （ 32.8 ＋ 3.8 ） respectively. The results were compared with that of routine one-step extraction method. CON- CLUSION： The three-step sequential protein extraction method has the higher protein extraction rate and the better protein dis- solubility as compared with one-step protein extraction method.
出处
《第四军医大学学报》
北大核心
2007年第21期1964-1966,共3页
Journal of the Fourth Military Medical University
关键词
总蛋白
蛋白质组
基质辅助激光解吸电离飞行时间质谱
提取
total protein
proteome
extraction
matrlx-assistedlaser desorption ionization-time of flight-mass spectra
