[Abstract] Nanofluidic proteomic immunoassay (NIA), consisting of isoelectric focusing followed by sensitive chemiluminescence detection, has the potential to quantitatively characterize protein post-translational modifications that cause shifts in isoelectric point (pI). This protocol details the NIA analysis of protein phosphorylation using AKT as an example.
This protocol can be used for two platforms, NanoPro 1000 and Peggy Sue, from ProteinSimple Company. NanoPro 1000 separates proteins based on charge. Peggy Sue separates proteins based on either charge or size. The platforms can ...[摘要] 纳米流体蛋白质组学免疫测定（NIA），由等电聚焦跟随敏感化学发光检测组成，具有定量表征导致等电点（pI）移位的蛋白质翻译后修饰的潜力。 该协议详细描述了使用AKT作为实例的蛋白质磷酸化的NIA分析。
此协议可用于ProteinSimple公司的两个平台NanoPro 1000和Peggy Sue。 NanoPro 1000基于电荷分离蛋白质。 Peggy Sue基于电荷或大小分离蛋白质。 平台可以一次分析高达96个样品，每个测定每裂解物少至25个细胞。 有关平台的详细信息，请访问ProteinSimple的网站（ http://www.proteinsimple.com ）。
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