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In Situ Hybridization and Immunostaining of Xenopus Brain.

Methods in molecular biology (Clifton, N.J.) (2019-09-26)
Kai-Li Liu, Xiu-Mei Wang, Zi-Long Li, Ying Liu, Rong-Qiao He
ABSTRACT

The dynamic expression pattern analysis provides the primary information of gene function. Differences of the RNA and/or protein location will provide valuable information for gene expression regulation. Generally, in situ hybridization (ISH) and immunohistochemistry (IHC) are two main techniques to visualize the locations of gene transcripts and protein products in situ, respectively. Here we describe the protocol for the whole brain dissection, the in situ hybridization, and the immunostaining of the developing Xenopus brain sections. Additionally, we point out the modification of in situ hybridization for microRNA expression detection.

MATERIALS
Product Number
Brand
Product Description

Roche
Anti-Digoxigenin-AP, Fab fragments, from sheep
Sigma-Aldrich
BCIP®/NBT Liquid Substrate System, ready to use solution