Anti seh antibody

The tissue fixation, paraffin embedding, section, and dewaxing were performed as previously described [49 (link)]. Antigen retrieval was performed by heating the sections in 0.01 M citrate buffer (pH 6.0) to 95 °C for 10 min. The immunohistochemistry staining was conducted using horseradish peroxidase (HRP)/3,3′-diaminobenzidine (DAB) Detection IHC kit (Abcam, Cambridge, UK) according to the manufacturer’s instructions. The anti-p21 antibody (Cell Signaling, Danvers, MA, USA, catalog # 64016), anti-Chop antibody (Cell Signaling, catalog # 2895), or anti-sEH antibody (Santa Cruz Biotechnology, Dallas, TX, USA, catalog # sc-166961) was used to probe the target protein in the tissue section. Sections were then counterstained with hematoxylin for 1 min. The staining intensity of p21, Chop, and sEH was analyzed by Image J software using IHC Toolbox.

Heart tissues or H9c2 cells were homogenized in 250 μ of homogenization buffer using an electronic stirrer. Protein concentration was determined with BCA kit (Biocolors, Shanghai, China). The process of western blot was used as previously described [32 (link)]. The following primary antibodies were used: polyclonal anti-sEH antibody (Santa Cruz Biotechnology) and anti-GAPDH antibody (Sigma) was used as an internal loading control.