Moens Cecilia
Division of Basic Science, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.
CSH Protoc. 2008 Aug 1;2008:pdb.prot5037. doi: 10.1101/pdb.prot5037.
INTRODUCTIONThis protocol describes procedures for detecting gene expression in zebrafish embryos in situ using labeled RNA probes. The labels (digoxigenin or fluorescein) are detected using alkaline phosphatase-conjugated antibodies that catalyze a chromogenic reaction, producing visible blue or red products. Two-color in situ hybridization can be performed by labeling with a combination of digoxigenin- and fluorescein-labeled probes and detecting the digoxigenin and fluorescein with sequential alkaline phosphatase reactions using different chromogenic substrates. In the method described here, the fluorescein-labeled probe is detected with the red substrate, but either probe can be detected with either substrate.
引言
本方案描述了使用标记RNA探针原位检测斑马鱼胚胎中基因表达的程序。使用催化显色反应的碱性磷酸酶偶联抗体检测标记（地高辛或荧光素），产生可见的蓝色或红色产物。双色原位杂交可通过用地高辛标记和荧光素标记的探针组合进行标记，并使用不同的显色底物通过连续碱性磷酸酶反应检测地高辛和荧光素。在此处描述的方法中，用红色底物检测荧光素标记的探针，但任一探针可用任一底物检测。
