Elimination of affinity reagent interference for the mass spectrometric detection of low-abundance proteins following immunoprecipitation.
Elimination of affinity reagent interference for the mass spectrometric detection of low-abundance proteins following immunoprecipitation.
复制标题
消除免疫沉淀后低丰度蛋白质质谱检测中亲和试剂的干扰。
DOI：
10.1021/pr070517a
复制
发表时间：
2007
影响因子：
4.4
通讯作者：
Sinai,AnthonyP
中科院分区：
文献类型：
--
作者：
Martin,AngelaM;Liu,Ting;Lynn,BertC;Sinai,AnthonyP
亲和试剂如免疫球蛋白存在于灵敏质谱分析的制剂中，可能会妨碍识别低丰度感兴趣的蛋白质。我们报告了一种在免疫沉淀中使用抗血清之前对其进行纯化和生物素化的方法，该方法允许通过捕获链霉亲和素有效地将其从蛋白质组样本中移除。这种方法同样可以推广到其他亲和试剂，如重组融合蛋白，以增强相互作用蛋白的鉴定。
The presence of affinity reagents such as immunoglobulin in preparations for sensitive mass spectrometry analyses can preclude the identification of low-abundance proteins of interest. We report a method whereby antisera are purified and biotinylated prior to use in immunoprecipitation that allows for its efficient removal from proteomic samples via streptavidin capture. This method can similarly be extended to other affinity reagents such as recombinant fusion proteins for enhanced identification of interacting proteins.