邦菲iTRAQ论坛

 找回密码
 立即注册

QQ登录

只需一步,快速开始

扫一扫,访问微社区

搜索
查看: 1513|回复: 0

尿液蛋白质组学实验方法建立

[复制链接]
发表于 2015-12-28 15:56:05 | 显示全部楼层 |阅读模式

了解更多iTRAQ信息,马上注册~

您需要 登录 才可以下载或查看,没有帐号?立即注册

x
Mol Cell Proteomics. 2010 Oct;9(10):2195-204. doi: 10.1074/mcp.M110.000992. Epub 2010 May 28.
Optimizing a proteomics platform for urine biomarker discovery.

Abstract
Biomarker discovery approaches in urine have been hindered by concerns for reproducibility and inadequate standardization of proteomics protocols. In this study, we describe an optimized quantitative proteomics strategy for urine biomarker discovery, which is applicable to fresh or long frozen samples. We used urine from healthy controls to standardize iTRAQ (isobaric tags for relative and absolute quantitation) for variation induced by protease inhibitors, starting protein and iTRAQ label quantities, protein extraction methods, and depletion of albumin and immunoglobulin G (IgG). We observed the following: (a) Absence of protease inhibitors did not affect the number or identity of the high confidence proteins. (b) Use of less than 20 μg of protein per sample led to a significant drop in the number of identified proteins. (c) Use of as little as a quarter unit of an iTRAQ label did not affect the number or identity of the identified proteins. (d) Protein extraction by methanol precipitation led to the highest protein yields and the most reproducible spectra. (e) Depletion of albumin and IgG did not increase the number of identified proteins or deepen the proteome coverage. Applying this optimized protocol to four pairs of long frozen urine samples from diabetic Pima Indians with or without nephropathy, we observed patterns suggesting segregation of cases and controls by iTRAQ spectra. We also identified several previously reported candidate biomarkers that showed trends toward differential expression, albeit not reaching statistical significance in this small sample set.


PMID: 20511394

免费全文链接http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2957724/


Mol Cell Proteomics-2010-IF7.251-方法学建立.pdf

471.31 KB, 下载次数: 1, 下载积分: 菲币 -2

邦菲生物——国内首推Orbitrap Fusion™蛋白质组快速鉴定服务
回复

使用道具 举报

您需要登录后才可以回帖 登录 | 立即注册

本版积分规则

北京邦菲生物科技有限公司招聘启事

手机版|Archiver|小黑屋|邦菲iTRAQ论坛 ( 京ICP备13049147号 )

GMT+8, 2018-8-15 20:59

Powered by Discuz! X3.2

© 2001-2013 Comsenz Inc.

快速回复 返回顶部 返回列表