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 language | English |
|---|---|
| Title of host publication | Methods in Molecular Biology |
| Publisher | Humana Press Inc. |
| Pages | 11-19 |
| Number of pages | 9 |
| Volume | 1489 |
| DOIs | |
| Publication status | Published - 2017 |
Publication series
| Name | Methods in Molecular Biology |
|---|---|
| Volume | 1489 |
| ISSN (Print) | 10643745 |
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Keywords
- Ccn2 gene expression
- Dig-labeled riboprobe
- Mouse embryo
- Whole-mount in situ hybridization
ASJC Scopus subject areas
- Molecular Biology
- Genetics
Cite this
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 proceeding › Chapter
}
TY - CHAP
T1 - Gene expression analysis of CCN proteins
T2 - Whole-mount in situ hybridization of Ccn2 in developing calcified tissues
AU - Yamaai, Tomoichiro
AU - Takigawa, Masaharu
PY - 2017
Y1 - 2017
N2 - 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.
AB - 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.
KW - Ccn2 gene expression
KW - Dig-labeled riboprobe
KW - Mouse embryo
KW - Whole-mount in situ hybridization
UR - http://www.scopus.com/inward/record.url?scp=84991710830&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84991710830&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-6430-7_2
DO - 10.1007/978-1-4939-6430-7_2
M3 - Chapter
C2 - 27734361
AN - SCOPUS:84991710830
VL - 1489
T3 - Methods in Molecular Biology
SP - 11
EP - 19
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -