Two-color detection of mRNA transcript localizations in fish and fly embryos using alkaline phosphatase and β-galactosidase conjugated antibodies
Two-color detection of mRNA transcript localizations in fish and fly embryos using alkaline phosphatase and β-galactosidase conjugated antibodies
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DOI：
10.1007/s004270050258
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发表时间：
1999-05-01
影响因子：
2.4
通讯作者：
Hauptmann, G
中科院分区：
文献类型：
--
作者：
Hauptmann, G
提出了一种快速、简化的双色原位杂交程序，用于同时检测完整斑马鱼和果蝇胚胎中两种不同的 mRNA。使用碱性磷酸酶和 β-半乳糖苷酶偶联抗体的混合物在单个孵育步骤中检测转录本分布。通过使用β-半乳糖苷酶底物以对比色可视化不同的转录本，该底物产生颜色沉淀（洋红色、蓝色），易于与标准碱性磷酸酶底物区分开。该方案之后可以进行标准免疫组织化学，以检测第三种颜色的第三种基因（在蛋白质水平）的表达。
A fast and simplified two-color in situ hybridization procedure for the simultaneous detection of two different mRNAs in whole-mounted zebrafish and Drosophila embryos is presented. Transcript distributions are detected in a single incubation step using a mixture of alkaline phosphatase and beta-galactosidase coupled antibodies. The different transcripts are visualized in contrasting colors by the use of beta-galactosidase substrates that develop color precipitates (magenta, blue) easily distinguishable from those of the standard alkaline phosphatase substrates. This protocol can be followed by standard immunohistochemistry to detect the expression of a third gene (at the protein level) in a third color.