蛋白质定量-实验方法

实验频道正文用户登录新用户注册蛋白质定量作者：佚名实验频道来源：中华基因网点击数：更新时间：2004-6-8 Quantitative Determination of Peptides by Sulfhydryl (-SH) Groups New (Contributed by David Van Horn, Dept. of Chemistry, UC Berkeley Greg Bulaj, Dept. of Biology, University of Utah)This is the standard “Ellman’s Test” for the determination of free thiols. It works well for small peptides and proteins synthesized using standard solid phase synthetic methods. Lowry Protein Assay (P.J. Hansen Lab, Dept. of Animal Sciences, University of Florida)The Lowry procedure is one of the most venerable and widely-used protein assays, being first described in 1951 [Lowry et al., J. Biol. Chem. 193: 265-275 (1951)]. Under alkaline conditions, copper complexes with protein. When folin phenol reagent (phospho-molybdic-phosphotungstic reagent) is added, the Folin-phenol reagent binds to the protein. Bound reagent is slowly reduced and changes color from yellow to blue. Lowry Protein Assay Lowry Assay for Protein Concentration (Folin-Ciocalteu Assay) Lowry Protein Assay (William H. Heidcamp) Bradford Protein Assay (P.J. Hansen Lab, Dept. of Animal Sciences, University of Florida)The Bradford protein assay is a simple procedure for determination of protein concentrations in solutions that depends upon the change in absorbance in Coomassie Blue G-250 upon binding of protein. Unlike many other assays, including the Lowry procedure, the Bradford assay is not susceptible to interference by a wide usr/localiety of chemicals present in samples. The notable exception is high concentrations of detergents. There is significant protein-to-protein usr/localiation in absorbance values obtained with the Bradford procedure and it is advisable to choose a protein standard that is likely to give absorbance values close to those for the protein samples of interest. The assay here is designed for use in microtiter plates. This is an easy assay format for those with access to multiple channel pipettors and microtiter plate spectrophotometers. Bradford Protein Assay (William H. Heidcamp) Biorad Protein Assay: Bradford Reagent (Suprya Jayadev) Biuret Protein Assay (William H. Heidcamp) Modified Lowry Protein Assay (Hancock Lab) The advantage of this assay over the Lowry assay from which it is derived, is the presence of SDS in reagent A. This allows one to perform protein assays in nearly any detergent (eg., when detergents are used in the purification of outer membrane proteins). In addition, SDS separates membrane proteins from contaminating membrane constituents and denatures the proteins allowing more reproducible results. Micro-Protein Assay (Jim Brown Lab)This micro-protein assay is not as accurate as other methods for assaying protein concentration, but it has the distinct advantage of requiring only trace amounts (less than 1ug) of your valuable protein. In many cases the accuracy (within a factor of 2) of the assay is more than sufficient. Another advantage of the assay is its relative ease and quickness Phosphoamino Acid Analysis of non-TCA Precipitable Proteins (Woodgett Lab) Protein Concentration of Laemmli Gel Samples (Woodgett Lab) 处理 SSI 文件时出错处理 SSI 文件时出错【评论】【进论坛看】【转入博客】实验频道录入：生物编辑责任编辑：生物编辑上一篇实验频道：肽段下一篇实验频道：细胞核提取【发表评论】【加入收藏】【告诉好友】【打印此文】【关闭窗口】相关文章最新热门推荐文章蛋白质提取与纯化技术2%多聚甲醛固定剂配方二则细胞学技术细胞培养的一般技术血小板的活化、荧光染色与流血小板活化检测方法细胞表面抗原染色的组织样本流式细胞仪PI染色死细胞方法流式细胞术分析血小板流式检测细胞周期 · 免疫细胞表面抗原分子CD家族对照表· 所有的看家基因(housekeeping gene· 生物发光与化学发光专题· 生物发光和化学发光在生物技术中的· 重组DNA的分离、克隆与测序实验手册· RNAi和基因沉默的历史回顾· RNAi原理图解· RNAi术语表· siRNA设计指南· Methods for DNA sequencing · 通用电气医疗集团简介· 生物发光与化学发光专题· 生物发光和化学发光在生物技术中的· RNA干扰技术获得新突破· 蛋白质特性与分离纯化技术的选择· PCR实验指导与常见问题分析· RNAi原理FLASH演示· Extrachromosomal elements, plasm· Promoter analysis by saturation · 重组DNA的分离、克隆与测序实验手册文章评论（只显示最新10条。评论内容只代表网友观点，与本站立场无关！）生物谷 | 网站介绍 | 网站地图 | 广告服务 | 帮助关于我们 | 广告服务 | 战略伙伴 | 留言| 帮助信息| 联系方式 | 客户投诉 | 友情链接 | 网站地图 | 网站声明 | Bioon English 上海北岸信息技术有限公司公司地址：上海龙吴路51号嘉源商务中心1#211室(200232) 生物谷网站 bioon.com © Copyright 2001 - 2006. All rights reserved. 备案号：沪ICP备05022939号
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