Hrp conjugated goat anti rat secondary antibody

Section and whole-mount RNA in situ hybridization and whole-mount X-gal or Salmon-gal staining to detect ß-galactosidase activity was performed as previously described (Anderson et al., 2004 (link); Kishigami et al., 2006 (link); Rojas et al., 2005 (link)). For Mef2c in situ hybridization, we used a 208-bp probe that corresponds to coding exon 2 as described elsewhere (Anderson et al., 2015 (link)). Lineage analysis using Etv2::Cre and Rosa26^LacZ/LacZ mice was performed as described previously (Verzi et al., 2005 (link)). For whole mount immunohistochemistry, rat anti- mouse CD31 (1:250, BD Biosciences, #553370) and horse radish peroxidase (HRP)-conjugated goat anti-rat secondary antibody (1:250, Abcam, ab7097) were used and detected with the DAB substrate kit (Vector, SK-4100), as described previously (Barnes et al., 2010 (link)). Immunofluorescence detection of ß-galactosidase, CD31, and myosin heavy chain (MF20) was performed as described previously (Schachterle et al., 2012 (link)). The following primary antibodies were used at a 1:100 dilution: rat anti- mouse CD31 (BD Biosciences, #553370), mouse anti-chicken MYH1E (DSHB, MF20), and chicken anti-ß-galactosidase (Abcam, ab9361). The following secondary antibodies were used at a 1:300 dilution: goat anti-rat AF488 (Abcam, ab150157), goat anti-mouse AF488 (Abcam, ab150113), and goat anti-chicken AF594 (Abcam, ab150172).