Phenol-based Total Protein Extraction from Lily Plant Tissues
从百合组织中提取苯酚标记总蛋白
发布: 2015年02月20日第5卷第4期 DOI: 10.21769/BioProtoc.1399 浏览次数: 12195
评审: Pablo Bolanos-VillegasArsalan DaudiAnonymous reviewer(s)
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参见作者原研究论文
The authors used this protocol in:
May 2014
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Abstract
The phenol-based total protein extraction method is unique in that water-soluble components such as polyphenolic compounds and nucleic acids can be easily removed. Thus, total protein is free from contaminants and allows for high quality two–dimensional gel electrophoresis. The phenol-based extraction of total protein was used in various lily organs and may likely apply to other plants whose content of polyphenolics is high (Note 1). An additional advantage of this extraction method is that nucleic acids can be easily removed and thus, avoid adverse effects of nucleic acids on protein resolution in the gel. This method is modified from that of Hurkman and Tanaka (1986).
Keywords: Protein extratction (蛋白质extratction)
Phenol (苯酚)
Plant (植物)
Materials and Reagents
- Sucrose (United States Biological, catalog number: 21938 )
- Tris base (United States Biological, catalog number: 75825 )
- HCl (fuming, at 37%) (Merck KGaA, catalog number: 100317 )
- EDTA (United States Biological, catalog number: 15699 )
- KCl (SERVA, catalog number: 26868 )
- 2-Mercaptoethanol (Sigma-Aldrich, catalog number: M6250 )
- Phenol (Merck KGaA, catalog number: 100206 )
- NH4OAc (Merck KGaA, catalog number: 101116 )
- 100% MeOH (J.T. Baker®, catalog number: 9093-03 )
- Urea (United States Biological, catalog number: 75826 )
- Triton X-100 (Sigma-Aldrich, catalog number: T9284 )
- K2CO3 (Merck KGaA, catalog number: 106683 )
- L-lysine (Sigma-Aldrich, catalog number: L5501 )
- DTT (Merck KGaA, catalog number: 111474 )
- Extraction buffer (see Recipes)
- Dissolving buffer (see Recipes)
- 0.1 M NH4OAc in 100% MeOH (see Recipes)
Equipment
- Ceramic mortar and pestles
- Beaker
- Stir bar
- Centrifuge (Eppendorf, model: 5424 )
- AVANTI® J-26 XP High-Performance centrifuge with a JA 25.50 fixed-angle rotor (Beckman Coulter)
- COREX® tube,DuPont Instruments (catalog number: 00152 )
- 1.5 ml Eppendorf tubes
- Fume hood
- Speed vacuum (Savant Systems LLC, model: SC110 )
- Mixer (Vortex Genie 2, Scientific Industries, Inc.)
- Stirrer (Corning)
Procedure
文章信息
版权信息
© 2015 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Chen, Y., Hsiao, Y. and Wang, C. (2015). Phenol-based Total Protein Extraction from Lily Plant Tissues. Bio-protocol 5(4): e1399. DOI: 10.21769/BioProtoc.1399.
Download Citation in RIS Format
分类
植物科学 > 植物生物化学 > 蛋白质 > 分离和纯化
植物科学 > 植物生理学 > 组织分析
生物化学 > 蛋白质 > 分离和纯化
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