Hrp conjugated goat anti rat igg

The following polyclonal (pAb) and monoclonal (mAb) primary antibodies used in the study were obtained from the indicated commercial sources: BAF155 rabbit pAb (1:20; Santa Cruz), BAF155 mouse mAb (1:100; Santa Cruz), BAF170 rabbit pAb (1:100; Bethyl), BAF170 rabbit pAb (1:100; Sigma), Olig2 rabbit pAb (1:200; Millipore), PDGFRα rat pAb (1:200; BD Bioscience), Sox9 rabbit pAb (1:100; Millipore), Sox10 Guinea pig pAb (1:100; a gift from Prof. Michael Wegner), Ki67 rabbit pAb (1:50; Novocastra), Ki67 mouse mAb (1:100; Novocastra), IdU mouse mAb (1:50; Becton Dickinson), Casp3 rabbit pAb (1:100; Cell Signaling).
Secondary antibodies used were horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (1:10,000; Covance), HRP-conjugated goat anti-mouse IgG (1:5,000; Covance), HRP-conjugated goat anti-rat IgG (1:10,000; Covance), and Alexa 488-, Alexa 568-, Alexa 594- and Alexa 647-conjugated IgG (various species, 1:400; Molecular Probes).

The following are the polyclonal (pAb) and monoclonal (mAb) primary antibodies used in this study: Pax6 rabbit pAb (1:200; Covance, Schwerte, Germany), H3K9Ac rabbit pAb (Abcam, Cambridge, UK), Sox2 goat (1:100; Santa Cruz, Dallas, TX, USA), Sox2 mouse mAb (1:100; R&D Systems, Minneapolis, MN, USA), Pax6 mouse mAb (1:100; Developmental Studies Hybridoma Bank, MA, USA), Tbr2 rabbit pAb (1:200; Abcam), H3ac rabbit pAb (Upstate, MA, USA), TNC rabbit pAb (Abcam), AP2γ mouse mAb (1:100; Abcam), PTPRZ1 rabbit pAb (Sigma, St. Louis, MO, USA), BAF155 mouse mAb (1:100; Santa Cruz), BAF155 rabbit pAb (1:20; Santa Cruz), Tbr1 rabbit pAb (1:300; Chemicon, St. Louis, MO, USA), HuCD mouse mAb (1:20; Invitrogen, Schwerte, Germany), Ctip2 rat pAb (1:200; Abcam), Cux1 rabbit pAb (1:100; Santa Cruz), Satb2 mouse mAb (1:200; Abcam), NeuN mouse mAb (Chemicon), Nestin mouse mAb (BD, Heidelberg, Germany), and Kat2a rabbit pAb (Abcam).
Secondary antibodies used were horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (1:10,000; Covance), HRP-conjugated goat anti-mouse IgG (1:5000; Covance), HRP-conjugated goat anti-rat IgG (1:10,000; Covance), and Alexa 488-, Alexa 568-, Alexa 594-, and Alexa 647-conjugated IgG (various species, 1:400; Molecular Probes, Schwerte, Germany). Antibodies were purchased from the indicated manufacturers in Germany.

To measure the protein expression level, the myocardial tissues and the cardiomyocytes were lysed by using RIPA lysis buffer (Thermo Fisher Scientific, Waltham, MA, USA). BCA protein assay kit (Thermo Fisher Scientific) was used to detect the protein concentration. Then, equal amounts of proteins (30 μg/lane) were separated by 10% SDS-PAGE and electro-transferred onto PVDF membranes (Thermo Fisher Scientific). Membranes were incubated with 5% non-fat milk for 2 h at room temperature, and then incubated with primary antibodies which were purchased from Abcam (Cambridge, UK) including rabbit monoclonal anti-DNMT3B (1:1000, ab79822), rabbit polyclonal anti-SOAT1 (1:1000, ab39327) and anti-Histone H3(1:2500, ab1791) and mouse monoclonal anti-GADPH (1:2000, ab9485) overnight at 4° C. And then the membranes were incubated with secondary antibody horseradish peroxidase (HRP)-conjugated goat anti-rat IgG (1:200, Covance, SMI-5040C) for 2 h, and GADPH was used as a loading control. The expression level of protein was analyzed by chemiluminescence and quantified using ImageJ software (National Institutes of Health, Bethesda, MA, USA) and E-Gel Imager (Thermo Fisher Scientific).