从百合组织中提取苯酚标记总蛋白

Yun-Chu Chen; Yu-Chun Hsiao; Co-Shine Wang

doi:10.21769/BioProtoc.1399

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Peer-reviewed

Phenol-based Total Protein Extraction from Lily Plant Tissues

从百合组织中提取苯酚标记总蛋白

YC Yun-Chu Chen

YH Yu-Chun Hsiao

CW Co-Shine Wang email

发布: 2015年02月20日第5卷第4期 DOI: 10.21769/BioProtoc.1399 浏览次数: 11277

评审: Pablo Bolanos-VillegasArsalan DaudiAnonymous reviewer(s)

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Cover of Journal of Experimental Botany, featuring study using the protocol.

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

English

中文翻译

文章信息

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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.