热搜: 微生物检验  分子生物学  检验基础  资格考试  医疗机构  实验室管理  有限公司  分子诊断学  荧光  胶体金 
 
 
当前位置: 首页 » 资料文档 » 分子识别与分离 »

[下载]Products for Affinity Purification affinity最新收录

PDF文档
  • 文件类型:PDF文档
  • 文件大小:15M
  • 更新日期:2015-02-25 19:43
  • 浏览次数:294   下载次数:954
进入下载
详细介绍
[下载]Products for Affinity Purification affinity最新收录Products for Affinity PurificationPublisher:PierceNumber of Pages:80Publication Date:2002Introduction:Various methods are used to enrich or purify a protein of interest from other proteins and components in a crude cell lysate or other sample. The most powerful of these methods is affinity purification, also called affinity chromatography, wherby the protein of interest is purified by virtue of its specific binding properties to an immobilized ligand. Pierce offers a number of immobilized protein or ligand products for affinity purification of antibodies, fusion-tagged proteins, biotinylated proteins and other proteins for which an affinity ligand is available.Affinity purification makes use of specific binding that occurs between molecules and is used extensively for the isolation of biological molecules. A single pass through an affinity column can achieve a 1,000- to 10,000-fold purification of a ligand from a crude mixture.From a single affinity purification step, it is possible to isolate a compound in a form pure enough to obtain a single band upon SDS-PAGE analysis.In affinity purification, a ligand is immobilized to a solid support. once immobilized, it specifically binds its partner under mild buffer conditions (often physiological conditions such as phosphate buffered saline). After binding to the partner molecule, the support is washed with additional buffer to remove unbound components of the sample. An elution buffer is added, disrupting the interaction between the ligand and its binding partner by pH extremes (low or high), high salt, detergents, chaotrophic agents or the removal of some factor required for the pair to bind. once released, the binding partner can be recovered from the support using additional elution buffer. The elution buffer can then be exchanged by dialysis or desalting into a more suitable buffer for storage or downstream analysis. Activated affinity support products and kits enable a researcher to immobilize nearly any type of ligand to purify its binding partner(s). For example, if a peptide antigen is used to immunize animals and produce antibodies, the same peptide may be immobilized to a gel support and used to affinity-purify the specific antibody from animal serum. Alternatively, if a specific antibody is available against a particular protein of interest, it can be immobilized to a support and used to affinity-purify the protein from crude cell lysate. Purification with respect to nearly any binding interaction can be made by this approach. Affinity purification products using either immobilized ligands or activated affinity support chemistries are available for use in several different formats. Most commonly, cross-linked beaded gel supports are used for gravity-flow column, spin-column or batch-scale purificationprocedures. Coated microplates are available for high-throughput screening applications, and magnetic particles are especially useful for affinity-based cell separation.Proteins and other macromolecules of interest can be purified from crude extracts or other complex mixtures using a variety of methods. Precipitation is perhaps the simplest method for separating one type of macromolecule from another. For example, nucleic acids can be precipitated and thereby purified from undesired molecules in solution using ethanol and proteins can be selecively precipitated in the presence of ammonium sulfate.Most purification methods involve some form of chromatography wherby molecules in solution (mobile phase) are separated based on differences in chemical or physical interaction with a stationary material (solid phase). Gel filtration (also called desalting, size-exclusion chromatography or SEC) uses a porous gel material to separate molecules based on size; large molecules are excluded from the internal spaces of the gel material while small molecules enter the resin pores, resulting in a longer path through the column. In ion-exchange chromatography, molecules are separated according to thestrength of their overall ionic interaction with a solid-phase material. By manipulating buffer conditions, molecules of greater or lesser ionic character can be bound to or dissociated from the solid-phase material.
 
[ 资料文档搜索 ]  [ 加入收藏 ]  [ 告诉好友 ]  [ 打印本文 ]  [ 关闭窗口 ]

下载地址



您还没有登录,请登录后查看详情
免费注册为会员后,您可以...
发布供求信息 推广企业产品
建立企业商铺 在线洽谈生意
还不是会员,立即免费注册
 


0条 [查看全部]  相关评论
 
更多»相关资料文档
推荐资料文档
本类下载排行
总下载排行
 
网站首页 | 关于我们 | 联系方式 | 使用协议 | 版权隐私 | 网站地图 | 排名推广 | 广告服务 | 网站留言 | RSS订阅 | 浙ICP备13009735号-1

京公网安备 11010602030045号