Goat anti mouse igg2b tritc

Whole-mount immunofluorescence was performed as previously described^{51 (link)} using the monoclonal antibodies MF20 and S46 (Developmental Studies Hybridoma Bank). The secondary reagents goat anti-mouse IgG1–FITC (fluorescein isothiocyanate) and goat anti-mouse IgG2b–TRITC (tetramethylrhodamine isothiocyanate) (SouthernBiotech) recognize S46 and MF20, respectively. MF20 or S46 positive signal were observed by confocal microscopy.

Whole-mount in situ hybridizations (WISH) were performed as described previously [30 (link)]. Digoxigenin-labeled probes for vegfaa, vegfab, kdrl, myl7, amhc, vmhc, nkx2.5 and hand2 were transcribed using T7 and SP6 RNA polymerase (Invitrogen, Carlsbad, CA, USA).
Whole-mount immunofluorescence was performed as described previously [30 (link)], using primary monoclonal antibodies against sarcomeric myosin heavy chain (MF20) and atrial myosin heavy chain (S46). MF20 and S46 were obtained from the Developmental Studies Hybridoma Bank (DSHB, Iowa City, IA, USA). The secondary antibodies used were goat anti-mouse IgG1 FITC and goat anti-mouse IgG2b TRITC (both 1:100; Southern Biotechnology Associates, Birmingham, AL, USA).