碱性磷酸酶
报告基因
胚胎干细胞
细胞培养
比色法
生物
计算生物学
细胞生物学
分子生物学
生物化学
化学
基因表达
基因
酶
遗传学
色谱法
作者
Meiyan Wang,Xinyi Wang,Haifeng Ye
出处
期刊:Bio-protocol
[Bio-Protocol]
日期:2023-01-01
卷期号:13 (3)
被引量:1
标识
DOI:10.21769/bioprotoc.4600
摘要
Secreted reporters have been demonstrated to be simple and useful tools for analyzing transcriptional regulation in mammalian cells. The distinctive feature of these assays is the ability to detect reporter gene expression in the culture supernatant without affecting the cell physiology or leading to cell lysis, which allows repeated experimentation and sampling of the culture medium using the same cell cultures. Secreted embryonic alkaline phosphatase (SEAP) is one of the most widely used reporter, which can be easily detected using colorimetry following incubation with a substrate, such as p-nitrophenol phosphate. In this report, we present detailed procedures for detection and quantification of the SEAP reporter. We believe that this step-by-step protocol can be easily used by researchers to monitor and measure molecular genetic events in a variety of mammalian cells due to its simplicity and ease of handling. Graphical abstract Schematic overview of the workflow described in this protocol.
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