Maize Endosperm Protein Extraction and Analysis
玉米胚乳蛋白的提取和分析
发布: 2013年07月20日第3卷第14期 DOI: 10.21769/BioProtoc.832 浏览次数: 14095
评审: Anonymous reviewer(s)
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参见作者原研究论文
The authors used this protocol in:
Aug 2012
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Abstract
Alcohol-solubility is the most characteristic feature of the zein proteins, the major storage protein in maize. Using sodium borate buffer system with added reducing agent, total proteins are isolated, and zein proteins are separated from non-zein proteins. The extraction effect is intuitive on a SDS-PAGE isolation system. In addition, a simple and rapid approach to extract zeins is introduced, taking full advantage of alcohol-solubility of zeins directly.
Keywords: Maize (玉米)
Seed (种子)
Zein (玉米醇溶蛋白)
Extraction (萃取)
Materials and Reagents
- Mature corn kernels
- Petroleum ether
- Ethanol
- β-mercaptoethanol
- Sodium dodecyl sulfonate (SDS)
- Urea
- Glycerol
- HCl
- Tris(Hydroxymethyl)aminomethane (Tris)
- Phenylmethanesulfonyl fluoride (PMSF)
- Methylene diacrylamide
- Bromophenol blue
- CHAPS (Sigma-Aldrich, catalog number: V900480-5G )
- Dithiothreitol (DTT)
- Liquid nitrogen
- ddH2O
- SDS-PAGE gel (15% separation)
- Coomassie brilliant blue (R250) staining buffer
- 30% acrylamide (see Recipes)
- Sodium borate buffer (see Recipes)
- 5x protein loading buffer (see Recipes)
- IPG solution (see Recipes)
Equipment
- A mortar and pestle
- Centrifuge (Eppendorf, model: 5415D )
- Shaker (Zhicheng, model: ZHWY-111C )
- Concentrator plus (Eppendorf, catalog number: 5305000.193 )
- Gel DocTM XR + System (Bio-Rad, catalog number: 170-8195 )
Procedure
文章信息
版权信息
© 2013 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Chen, X., Yao, D. and Song, R. (2013). Maize Endosperm Protein Extraction and Analysis. Bio-protocol 3(14): e832. DOI: 10.21769/BioProtoc.832.
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分类
植物科学 > 植物生物化学 > 蛋白质 > 分离和纯化
生物化学 > 蛋白质 > 分离和纯化
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