期刊文献+
任意字段
题名或关键词
题名
关键词
文摘
作者
第一作者
机构
刊名
分类号
参考文献
作者简介
基金资助
栏目信息

两种方法提取脑组织总蛋白的色谱串联质谱分析 被引量：2

Chromatography-tandem mass spectrometry analysis of total protein in brain tissue by two extraction methods
下载PDF
导出
摘要 目的通过对提取大鼠脑组织总蛋白不同方法的研究,建立提取脑组织总蛋白的标准方法。方法在本实验中分别用总蛋白提取试剂盒和自制裂解液提取脑组织总蛋白,测量蛋白浓度。用体积排阻色谱(SEC)分离后分别进行基质辅助激光电离飞行时间质谱(MALDI-TOF-MS)做蛋白全谱。结果总蛋白提取试剂盒和自制裂解液总蛋白提取量分别为:(5.6±0.2)mg、(6.7±0.5)mg,MALDI-TOF-MS蛋白全谱的谱峰数分别为(304.2±22.3)和(331.6±30.5)个。结论自制裂解液与常规的总蛋白提取试剂盒相比具有蛋白提取率高、蛋白溶解性好等优点。 Objective To establish the standard method of total protein extraction from brain tissue. Methods The total protein extraction kit and self-made lysate were used to extract the total protein from brain tissues, and the protein levels were measured. The proteins were isolated using size exclusion chromatography （SEC） and detected by matrix-assisted laser desorption/ionization-time of flight-mass spectra （MALDI-TOF-MS）. Results The extraction amount of total protein by total protein extraction kit and self-made lysate were （5.6 ±0.2） mg and （6.7 ±0.5） mg, respectively. The MALDI-TOF-MS spectra peak was （304. 2 ±22. 3） and （331.6 ±30. 5）. Conclusion The self- made lysate extraction method has the advantage of higher protein extraction rate and the better protein dissolubility compared with total protein extraction kit extraction method.
作者 党军强 李学文 刘卫平 李丙所 周红兵 陈勇
机构地区 第四军医大学西京医院肝胆外科 定西市人民医院神经外科 第四军医大学西京医院神经外科
出处 《中华神经外科疾病研究杂志》 CAS 2009年第5期413-416,共4页 Chinese Journal of Neurosurgical Disease Research
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 裂解液 Total protein Proteome Matrix-assisted laser desorption/ionization-time of flight-tress spectra Lysate
分类号 R735.9 [医药卫生—肿瘤]
  • 相关文献

参考文献11

  • 1Jemal A, Siegel R, Ward E, et al. Cancer statistics, 2006 [ J ]. CA Cancer J Clin, 2006, 56(2): 106- 130.
  • 2DeAngelis LM. Brain tumors [J]. N Engl J Med, 2001,344(2) : 114 - 123.
  • 3Samoylova TI, Morrison NE, Cox NR. Molecular markers of glial tumors: current targeting strategies [ J]. Curr Med Chem, 2003, 10 (10) : 831 -843.
  • 4Sinha A, Singh C, Parmar D, et al. Proteomics in clinical interventions: achievements and limitations in biomarker development [J]. LifeSci, 2007, 80(15): 1345-1354.
  • 5Huang HQ, Xiao ZQ, Lin QM, et al. Characteristics of structure, composition, mass spectra, and iron release from the ferritin of shark liver ( Sphyrna zygaena ) [J]. Biophys Chem, 2004, 111 (3) : 213 - 222.
  • 6张久权,冯华.蛋白质组学及其在胶质细胞瘤研究中的进展[J].中华神经外科疾病研究杂志,2008,7(4):381-383. 被引量：2
  • 7Gao MX, Hong J, Yang PY, et al. Chromatographic prefractionation prior to two-dimensional electrophoresis and mass spectrometry identifies: Application to the complex proteome analysis in rat liver [J]. Analytica Chimica Acta, 2005, 553( 1 ) : 83 -92.
  • 8Fazal MA, Palmer VR, Dovichi NJ, el al. Analysis of differcntial detergent fractions of an ART-20 cellular homogenate using one-and two- dimensional capillary eleetrophorcsis [ J ]. J Chromatogr A, 2006, 1130(2) : 182 -189.
  • 9Kashino Y, Harayama T, Pakrasi HB, et al. Preparation of membrane proteins for analysis by two-dimensional gel electrophoresis[ J ]. J Chromatogr B Analyt Technol Biomed Life Sci, 2007, 849 ( 1 - 2 ) : 282 - 292.
  • 10Maddalo G, Petrucci F, lezzi M, et al. Analytical assessment of MALDI-TOF Imaging Mass Spectrometry on thin histological samples. An insight in protcomc investigation [J]. Clin Chim Acta, 2005, 357 (2) : 210 -218.

二级参考文献20

  • 1Tyers M, Mann M. From genomics to proteomics [J]. Nature, 2003, 422(6928) : 193 - 197.
  • 2Wasinger VC, Cordwell SJ, Cerpa-Poljak A, et al. Progress with geneproduct mapping of the Mollicutes: Mycoplasma genitalium [ J ]. Electrophoresis, 1995, 16(7): 1090- 1094.
  • 3Wilkins MR, Appel RD. Ten Years of the Proteome [A]. In: Wilkins MR, Appel RD, Williams KL, et al. (eds.). Proteome Research [ MJ. 2nd ed. Verlag Berlin Heidelberg: Springer, 2007:1 - 14.
  • 4Aebersold R, Mann M. Mass spectrometry-based proteomics [ J ]. Nature, 2003, 422 (6928) : 198 - 207.
  • 5Boguski MS, McIntosh MW. Biomedical informatics for proteomics [J]. Nature, 2003, 422(6928) : 233 -237.
  • 6Cravatt BF, Simon GM, Yates JR 3rd. The biological impact of massspectrometry-based proteomics [ J ]. Nature, 2007, 450 ( 7172 ) : 991 - 1000.
  • 7Domon B, Aebersold R. Mass spectrometry and protein analysis [ J]. Science, 2006, 312(5771 ): 212-217.
  • 8Hanash S. Disease proteomics [ J]. Nature, 2003,422(6928) : 226 - 232.
  • 9de Groot JF, Gilbert MR. New molecular targets in malignant gliomas [J]. Curr Opin Neurol, 2007, 20(6) : 712 -718.
  • 10Dalton WS, Friend SH. Cancer biomarkers-an invitation to the table [J]. Science, 2006, 312(5777): 1165-1168.

共引文献1

同被引文献31

  • 1高月,马增春,张伯礼.中药大品种二次开发的安全性关注及再评价意义[J].中国中药杂志,2012,37(1):1-4. 被引量：14
  • 2李晓光,高勤,翁文,章佩芬,肖飞,罗焕敏.五味子有效部位及其药理作用研究进展[J].中药材,2005,28(2):156-159. 被引量：127
  • 3蔡静.蛋白质丙酮沉淀对2-DE图谱的影响[J].第三军医大学学报,2006,28(3):270-271. 被引量：7
  • 4Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding[J]. Anal Biochem, 1976,72(1~2):248~254.
  • 5Grg A, Boguth G, Obermaier C, et al. Two-dimensional polyacrylamide gel electrophoresis with immobilized pH gradients in the first dimensin (IPG-Dalt): The state of the art and the controversy of vertical versus horizontal systems[J]. Electrophoresis, 1995,16(1):1079~1086.
  • 6Grg A, Weiss W, Dunn M J. Current two-dimensional electrophoresis technology for proteomics[J]. Proteomics, 2004, 4(12): 3665~3685.
  • 7Pathak N N, Pattanaik A K, Patra R C, et al. Mineral composition of antlers of three deer species reared in captivity[J]. Small Ruminant Res, 2001, 42(1): 61~65.
  • 8Patrick H, O’Farrell. High resolution two-dimensional electrophoresis of protein [J].The Journal of Biological Chemistry, 1975,250(10): 4007~4021.
  • 9Rabilloud T. Two-dimensional gel electrophoresis in proteomics: old, old fashioned, but it still climbs up the mountains[J]. Proteomics,2002,2(1):3~10. 3.0.CO;2-R target="_blank">.
  • 10Shaw M M, Riederer B M. Sample preparation for two-dimensional gel electrophoresis[J]. Proteomics, 2003,3(8):1408~1417.

引证文献2

二级引证文献9

中华神经外科疾病研究杂志
2009年 第5期

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部