Goat anti chicken igy alexa fluor 647

Immunostaining was carried out as described previously [4 (link), 21 (link)]. The following primary antibodies were used: mouse anti-FLAG® M2 (Sigma-Aldrich, cat. no. F1804; 1:200), rabbit anti-cardiac troponin I (Santa Cruz Biotechnology, cat. no. sc-15368; 1:100), mouse anti-cardiac troponin T (Thermo Scientific, cat. no. MS-295-P; 1:200), rabbit anti-Nanog (Cell Signaling Technology, cat. no. #4903; 1:50), rabbit anti-TRA-1–60 (Abcam, cat. no. ab174813; 1:100), chicken anti-beta III tubulin (Abcam, cat. no. ab41489; 1:1000), rabbit anti-SATB2 antibody (Abcam, cat. no. ab34735; 1:500), rabbit anti-TBR1 antibody (Abcam, cat. no. ab31940; 1:200), and chicken anti-tyrosine hydroxylase antibody (Abcam, cat. no. ab76442; 1:1000). Secondary antibodies used were as follows: goat anti-mouse IgG-Alexa Fluor® 546 (Life Technologies, cat. no. A-11030; 1:400), goat anti-rabbit IgG-Alexa Fluor® 647 (Life Technologies, cat. no. A-20991; 1:400), goat anti-chicken IgY-Alexa Fluor® 647 (Life Technologies, cat. no. A-21449; 1:400), and goat anti-chicken IgY-Alexa Fluor® 488 (Abcam, cat. no. ab150169; 1:400).
Hoechst 33342 (Life Technologies, cat. no. H3570; 1:10000) was used to counterstain nuclei. The stained cells and tissue sections were visualized using a confocal microscope (Olympus, FV1000) and fluorescence microscope (Keyence, BZ-X710).