Gene expression analysis of CCN proteins: Whole-mount in situ hybridization of Ccn2 in developing calcified tissues

Tomoichiro Yamaai, Masaharu Takigawa

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

A procedure for whole-mount in situ hybridization developed for detecting gene expression of Ccn2 in developing calcified tissues of mouse embryos is presented. In this method, embryos are hybridized with Dig-labeled riboprobes, and the riboprobes are detected by use of the alkaline-phosphatase reaction in the presence of a 4-nitro-blue tetrazolium chloride and 5-bromo-4-chloro-3-indolyl-phosphate (NBT + BCIP) mixture. Obvious detection of positive signals for Ccn2 in the cartilage of developing phalanges indicates that this method can be applied to gene expression analysis of other Ccn genes in developing calcified tissues.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages11-19
Number of pages9
Volume1489
DOIs
Publication statusPublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1489
ISSN (Print)10643745

Fingerprint

In Situ Hybridization
Gene Expression
Embryonic Structures
Proteins
Cartilage
Alkaline Phosphatase
Chlorides
Genes
5-bromo-4-chloro-3-indoxyl phosphate
4-anisyltetrazolium blue

Keywords

  • Ccn2 gene expression
  • Dig-labeled riboprobe
  • Mouse embryo
  • Whole-mount in situ hybridization

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Yamaai, T., & Takigawa, M. (2017). Gene expression analysis of CCN proteins: Whole-mount in situ hybridization of Ccn2 in developing calcified tissues. In Methods in Molecular Biology (Vol. 1489, pp. 11-19). (Methods in Molecular Biology; Vol. 1489). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-6430-7_2

Gene expression analysis of CCN proteins : Whole-mount in situ hybridization of Ccn2 in developing calcified tissues. / Yamaai, Tomoichiro; Takigawa, Masaharu.

Methods in Molecular Biology. Vol. 1489 Humana Press Inc., 2017. p. 11-19 (Methods in Molecular Biology; Vol. 1489).

Research output: Chapter in Book/Report/Conference proceedingChapter

Yamaai, T & Takigawa, M 2017, Gene expression analysis of CCN proteins: Whole-mount in situ hybridization of Ccn2 in developing calcified tissues. in Methods in Molecular Biology. vol. 1489, Methods in Molecular Biology, vol. 1489, Humana Press Inc., pp. 11-19. https://doi.org/10.1007/978-1-4939-6430-7_2
Yamaai T, Takigawa M. Gene expression analysis of CCN proteins: Whole-mount in situ hybridization of Ccn2 in developing calcified tissues. In Methods in Molecular Biology. Vol. 1489. Humana Press Inc. 2017. p. 11-19. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-6430-7_2
Yamaai, Tomoichiro ; Takigawa, Masaharu. / Gene expression analysis of CCN proteins : Whole-mount in situ hybridization of Ccn2 in developing calcified tissues. Methods in Molecular Biology. Vol. 1489 Humana Press Inc., 2017. pp. 11-19 (Methods in Molecular Biology).
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