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His标签蛋白检测试剂盒—HisDetector™ Western Blot Kits

27 March 2009 文章作者:房浩霞 有1,186人对该文章感兴趣 24 评论

在重组蛋白的纯化过程中,常常利用各种亲和标签来纯化所表达的融合蛋白,以满足各种下游实验的需求。在众多标签中,His标签是专门设计用于重组蛋白质的纯化,与其他标签相比有很多明显优势,是目前用于纯化的融合标签中使用最为广泛的一种。His标签专为重组蛋白纯化设计,与其它标签相比具有很多优势,如表达方便,分子小基本不影响蛋白的活性等。

通常对His标签重组蛋白的检测都会用特异的一抗或标签抗体来进行。使用特异一抗检测较为灵敏,但是当His标签的重组蛋白非常多时,购买相应的一抗做检测,就其各自的成本及实验优化时间来说都不是最优的方案。常规的标签抗体步骤是使用抗His标签的一抗,然后用酶标的二抗结合,之后使用相应的显色试剂或底物完成Western Blot检测。以上常规的检测方案都需要涉及到完整的一抗、二抗及底物显色步骤,在时间及经济节约上都有一定的缺陷。

His标签蛋白检测试剂盒—HisDetector™ Western Blot Kits

KPL利用Ni-NTA可与His组氨酸残基特异性结合的特性,开发了一系列针对His标签蛋白的检测产品以替代现有的His标签抗体及相应的检测方案。在Ni-NTA的镍上标记有HRP或AP,可直接催化底物显色或发光。采用这样的酶标Ni-NTA可以直接与His标签蛋白结合,然后直接显色,省去了一抗及二抗结合洗脱的时间,而且大大提高了检测灵敏度,为His标签蛋白检测提供了一个最优的分析方案。

与传统的His标签抗体检测手段相比,HisDetector™ 具有以下的优势:

  • HRP/AP-Ni与His结合,不再需要标签抗体和二抗;
  • 可检测4~10个组氨酸不等的His标签且不区分N-或C-端;
  • 操作简便更省时,全过程可节省2小时;
  • 镍与组氨酸的结合,特异性更强;
  • 高灵敏度,可检测pg级蛋白。

HisDetector™ Western Blot Kit试剂盒分AP底物显色试剂盒、HRP底物显色及化学发光试剂盒共三种,试剂盒内提供了酶标记的HisDetector™ Ni-NTA以及WB所需的封闭液、洗脱液以及底物。还有单独的酶标HisDetector™ Ni-NTA提供,分别为HRP及AP标记的Ni-NTA。

产品名称 货号 产品说明(点击查看说明书)
HisDetector Western Blot Kit, AP Colorimetric 25-00-01 底物显色,40次WB检测
HisDetector Western Blot Kit, HRP Chemiluminescent 24-00-02 化学发光,40次WB检测
HisDetector Western Blot Kit, HRP Colorimetric 24-00-01 底物显色,40次WB检测
产品名称 货号 产品说明(点击查看说明书)
HisDetector Nickel-AP 25-01-01 1mL Nickel-AP
HisDetector Nickel-HRP 24-01-01 0.1mL Nickel-HRP

作为美国最早实现亲和素纯化二抗商业化的生物公司,同时也是世界上最大的二抗和底物显色系统的生产商。 KPL一直提供高质量的用于分子生物学、免疫学、细胞生物学和体外诊断试剂及用于蛋白和核酸检测的试剂盒。针对于His重组蛋白的检测,KPL有HRP及AP标记的镍-NTA结合物,还有完整的HisDetector 标签蛋白Wester Blot检测试剂盒提供。另外还有多种化学发光和显色底物及相关产品,以及全系列针对不同种属的各种标记的现货二抗。如果您对KPL的产品感兴趣,请致电010-88597838/9569 /8795/4237到西美杰科技有限公司免费索取相关产品资料,或者问询您所感兴趣的产品类型。

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