Chapter title |
Gene Expression Analysis of CCN Proteins: Whole-Mount In Situ Hybridization of Ccn2 in Developing Calcified Tissues.
|
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Chapter number | 2 |
Book title |
CCN Proteins
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Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-6430-7_2 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6428-4, 978-1-4939-6430-7
|
Authors |
Tomoichiro Yamaai, Masaharu Takigawa D.D.S., Ph.D., Masaharu Takigawa |
Editors |
Masaharu Takigawa |
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. |
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