条带转移(Band Shift)-实验方法

实验频道正文用户登录新用户注册条带转移(Band Shift) 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 Or gel mobility shift assay, gel shift assay, gel retardation, electrophoretic mobility shift assay (EMSA) EMSA Using Oligos (Mike A. Dyer)Anneal two complementary oligos to make ds oligos as the probe for EMSA assay. Gel shift Assay (Hahn Lab) Band shifting (Promega)The gel shift assay is performed by incubating a purified protein, or a complex mixture of proteins (such as nuclear or cell extract preparations), with a 32P end-labeled DNA fragment containing the putative protein binding site. The reaction products are then analyzed on a nondenaturing polyacrylamide gel. The specificity of the DNA-binding protein for the putative binding site is established by competition experiments using DNA fragments or oligonucleotides containing a binding site for the protein of interest, or other unrelated DNA sequences. Bank Shift Assay for Kd Determinationi (Neris Lab, Institute of Pharmaceutical Sciences ETHZ)The procedure is for the determination of affinity constants and kinetic dissociation constants by band-shift assay refers to an ideal antibody fragment (e.g., a scFv or an Fab fragment) binding to a well-behaved protein antigen (pure, of well-defined oligomeric state, migrating as a single band in non-denaturing gel electrophoresis. As mentioned in the introduction, either the antibody or the antigen has to be labeled in these assays. Fluorescent Band Shift Assay (Amersham)The band shift assay is a useful tool for identifying protein-DNA interactions and can be used to determine the affinity, abundance, binding constants, and binding specificity of DNA-binding proteins. The gel shift assay is performed by annealing two labeled oligonucleotides that contain the test binding sequence, then incubating the duplex with the binding protein. The mixture is then separated on a nondenaturing polyacrylamide gel. Duplexes that are bound by protein migrate more slowly than unbound duplexes and appear as bands that are shifted relative to the bands from the unbound duplexes.This protocol using fluorescein end-labeled oligonucleotides as probe for DNA-protein binding. Fluorescence imaging provides a rapid, sensitive, and quantitative alternative to radioactivity for performing band shift. 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： M13噬菌体下一篇实验频道：足迹法(Footprinting) 【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免疫组化染色过程中存在的问常用核酸和氨基酸代码动物模型的设计原则和注意事PCR常见问题总汇如何确定RNA质量的经验谈PCR引物设计的11条黄金法则蛋白纯化经验指南TA克隆常见问题分析及其解决免疫组化实验过程中的要点和核酸的抽提---mRNA的分离技巧 · 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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