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学科主题物理化学
An activity-maintaining sequential protein extraction method for bioactive assay and proteome analysis of velvet antlers
Sui, Zhigang1; Yuan, Huiming1; Liang, Zhen1; Zhao, Qun1,2; Wu, Qi1,2; Xia, Simin1,2; Zhang, Lihua1; Huo, Yushu1; Zhang, Yukui1; Zhang LH(张丽华)
关键词Proteome Profiling Velvet Antlers Sequential Protein Extraction Method Activity Assay Huvec Cells
刊名TALANTA
2013-03-30
DOI10.1016/j.talanta.2013.01.015
107页:189-194
收录类别SCI
合作性质
文章类型Article
部门归属18
项目归属1810
产权排名待补充
WOS标题词Science & Technology ; Physical Sciences
资助者1,1 ; 1,1 ; 1,1 ; 1,1
类目[WOS]Chemistry, Analytical
研究领域[WOS]Chemistry
关键词[WOS]RED DEER ANTLER ; EXPRESSION ; PROLIFERATION ; REGENERATION ; GENOME ; GROWTH ; TISSUE ; VEGF
英文摘要The exceptional growth rate of velvet antler makes it a valuable model for studying the development of tissues, such as blood vessels, cartilage and bone. Meanwhile, investigating the activities of extracted proteins from velvet antlers promisingly leads to the discovery of new active factors which regulate the development of above-mentioned tissue types. In this study, a novel sequential protein extraction method was developed for proteome profiling and bioactivity study of velvet antlers. Herein, four antler growing tips were pooled to create a proportional pooled sample, and three aliquots of which were extracted in parallel using the developed extraction method. For each sample, proteins were extracted sequentially by saline solvent (0.15 M sodium chloride, pH 7.0), mild acid buffer (0.15 M acetate buffer, pH 4.0) and mild alkaline buffer (0.15 M glycine-sodium hydroxide buffer, pH 10.0) with good bio-compatibility to prevent proteins denaturation. Then STD lysis buffer, containing 4% SDS, 0.1 M Tris-HCl and 0.1 M DTT, was used to extract hydrophobic proteins. The tryptic digest of each fraction was analyzed by nanoRPLC-ESI-MS/MS in triplicates, with false discovery rate for peptide identification adjusted to 1% to create filtered protein group list. In total, 1423 protein groups were identified, which expanded up to 3 times of the previous published dataset The relative standard deviation of identified peptide and protein group number for all analyses indicated the good reproducibility of the developed sequential protein extraction method. Additionally, proteins extracted by acid buffer and alkaline buffer showed obvious promoting effect on the proliferation of human umbilical vein endothelial cells. All these results demonstrate that the developed sequential extraction method is efficient for the comprehensive proteome analysis and activity investigation of velvet antlers. (C) 2013 Elsevier B.V. All rights reserved.
语种英语
资助者1,1 ; 1,1 ; 1,1 ; 1,1
原文出处查看原文
WOS记录号WOS:000319088700028
引用统计
被引频次:10[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/119107
专题中国科学院大连化学物理研究所
通讯作者Zhang LH(张丽华)
作者单位1.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100093, Peoples R China
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Sui, Zhigang,Yuan, Huiming,Liang, Zhen,et al. An activity-maintaining sequential protein extraction method for bioactive assay and proteome analysis of velvet antlers[J]. TALANTA,2013,107:189-194.
APA Sui, Zhigang.,Yuan, Huiming.,Liang, Zhen.,Zhao, Qun.,Wu, Qi.,...&张丽华.(2013).An activity-maintaining sequential protein extraction method for bioactive assay and proteome analysis of velvet antlers.TALANTA,107,189-194.
MLA Sui, Zhigang,et al."An activity-maintaining sequential protein extraction method for bioactive assay and proteome analysis of velvet antlers".TALANTA 107(2013):189-194.
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