Embryonic Development: In Situ Hybridization and Antibody Staining

In situ hybridization and antibody staining was carried out as described previously [65 (link)–67 (link)]. In situ TUNEL assay was performed by using Promega terminal deoxynucleotidyl transferase (M828A) according to the manufacturer’s protocol. Following primary and secondary antibodies were used in this study: anti-Islet1/2 (Developmental Studies Hybridoma Bank 39.4D5, 1:100 for whole-mount, 1:250 for cryo-sections), anti-BrdU (Beckton-Dickinson, 1:250) and Alexa 546 goat anti-mouse IgG (Invitrogen A-11003, 1:50 for whole-mount, 1:250 for cryo-sections). Cryo-sectioning and BrdU labeling were carried out as described previously [5 (link)]. Whole-mount stained embryos were sectioned except for Figs. 1, 7 and S4 where anti-Islet1/2 staining was performed on sections using standard whole mount protocols.

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Kantarci H., Edlund R.K., Groves A.K, & Riley B.B. (2015). Tfap2a Promotes Specification and Maturation of Neurons in the Inner Ear through Modulation of Bmp, Fgf and Notch Signaling. PLoS Genetics, 11(3), e1005037.

Publication 2015

anti-Islet1/2 anti-BrdU

Anti igg Antibodies Antibody Assay Brdu Embryos Figs Goat Hybridoma In situ hybridization Mouse Promega Terminal deoxynucleotidyl transferase Tunel

Corresponding Organization : Baylor College of Medicine

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Variable analysis

independent variables

In situ hybridization
Antibody staining
TUNEL assay

dependent variables

Expression of Islet1/2
BrdU labeling

control variables

Protocols used for in situ hybridization and antibody staining as described previously [65 (link)–67 (link)]
Promega terminal deoxynucleotidyl transferase (M828A) used for TUNEL assay according to the manufacturer's protocol
Cryo-sectioning and BrdU labeling carried out as described previously [5 (link)]

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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