Comparison of Protein Extraction Methods for the Semen Seu Fructus Physalis

LI Xiao-lin; SHAO Ai-juan; CHEN Min; HUANG Lu-qi~

doi:10.13422/j.cnki.syfjx.2010.01.009

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Comparison of Protein Extraction Methods for the Semen Seu Fructus Physalis

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- Comparison of Protein Extraction Methods for the Semen Seu Fructus Physalis
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 16, Issue 1, Pages: 14-17(2010)
- 作者机构：
  
  中国中医科学院中药研究所
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.2010.01.009
  CLC： R284.1
- Published：2010
- 稿件说明：
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[1]李晓琳,邵爱娟,陈敏,黄璐琦.酸浆种子蛋白提取方法的比较研究[J].中国实验方剂学杂志,2010,16(01):14-17.

LI Xiao-lin, SHAO Ai-juan, CHEN Min, et al. Comparison of Protein Extraction Methods for the Semen Seu Fructus Physalis[J]. Chinese journal of experimental traditional medical formulae, 2010, 16(1): 14-17.
[1]李晓琳,邵爱娟,陈敏,黄璐琦.酸浆种子蛋白提取方法的比较研究[J].中国实验方剂学杂志,2010,16(01):14-17. DOI： 10.13422/j.cnki.syfjx.2010.01.009.

LI Xiao-lin, SHAO Ai-juan, CHEN Min, et al. Comparison of Protein Extraction Methods for the Semen Seu Fructus Physalis[J]. Chinese journal of experimental traditional medical formulae, 2010, 16(1): 14-17. DOI： 10.13422/j.cnki.syfjx.2010.01.009.

摘要

目的:建立适用于酸浆种子的SDS-PAGE和2-D PAGE蛋白样品提取方法。方法:比较直接提取法、TCA-丙酮法、Tris-丙酮法、饱和酚法、改良饱和酚法等7种提取方法所获酸浆种子总蛋白含量以及SDS-PAGE电泳的谱带数目、强度和电泳分辨率等方面的差异。结果:HEPES buffer直接提取法提取蛋白效率较高

杂质少

电泳谱带清晰

分辨率高;PVP buffer和Lysis buffer A直接提取法次之

但杂质较多;TCA-丙酮法、Tris-丙酮法、饱和酚法和改良的饱和酚法差别不大

蛋白纯度较高

但提取效率低

谱带数目少;此外

Lysis buffer A法、丙酮法和酚法易产生高丰度蛋白的干扰。结论:直接提取法中的HEPES buffer法最好

优于丙酮法、酚法和其它直接提取法。

Abstract

Objective:To compare seven protein extraction protocols（PVP buffer

HEPES buffer

Lysis buffer A extraction and TCA/acetone

tris-base/acetone

phenol/ammonium acetate

acetone/phenol/ammonium acetate precipitation） and establish a suitable one for total proteins extraction from Semen Seu Fructus Physalis.Methods:The protein yield

protein purity and SDS-PAGE patterns was determined and analyzed.Results:HEPES buffer extraction produce the purer sample

much more clear bands and highest resolution.PVP buffer and Lysis buffer A extraction had most impurities.TCA/acetone

tris-base/acetone

phenol/ammonium acetate

acetone/phenol/ ammonium acetate precipitation methods had the purest sample

but protein yield and band numbers were lowest.Besides

the methods employing Lysis buffer was easy to produce high-abundance proteins and interfered in the electrophoresis resolution.Conclusion:HEPES buffer extraction was the best sample preparation protocol for SDS-PAGE and 2D PAGE.

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