Caspase 3 antibody

Hepatic and renal specimens were cut into slices, dehydrated, and embedded in paraffin for histological analysis. Slices were then cut into 3 μm thick to be stained with hematoxylin and eosin (H&E), then visualized under an optical microscope.
Slices were embedded in paraffin, deparaffinized, and remoistened for immunohistochemical analysis. They were then cleaned in PBS and immersed for 15 min in 2 percent H2O2 to impede peroxidase activity. Bovine serum albumin (5%) was used to block non-specific binding sites. Bcl-2 Antibody [(C-2): sc-7382], Caspase-3 Antibody [(9CSP01): sc-81,663], and polyclonal antibodies (Santa Cruz Biotechnology, USA) were employed to coat the kidney and liver tissue specimen slides before being diluted to 1:500 and added. The slides were then kept at 4 °C for overnight incubation. A biotin-conjugated secondary antibody (catalog # sc-2040) was implemented to the slides after three PBS rinses. These were created with 3,3-diaminobezidine tetrahydrochloride, and hematoxylin was used as a counterstain [44 (link)]. The relative proportions of immune reactive cells for caspase-3 and Bcl2 measured by the ratio of positively stained cells to the total number of examined cells. Three slides from six rats in each group were the subjects of ANOVA tests to determine their significance.

For immunohistochemical (IHC) staining, primary antibodies of caspase-3 antibody (Santa Cruz Biotechnology, Dallas, TX: 1/100 and Cell Signaling Technology Inc., Danvers, MA) were used. Use was made of an automatic IHC device (Leica Bond-Max, Australia) to stain the sections. Having been processed in IHC device, the sections were dehydrated through a graded series of ethanol to xylene and closed (Entellan, Merck Millipore, Germany). For IHC operation, sections with a thickness of 4 μm were formed on a positively charged microscope slides were formed of the tissue samples in 10% formalin solution. The results of the analysis performed with Olympus BX51 microscope were assessed with caspase-3 staining of the tissues.

Picroside II was purchased from Shanghai Standard Biotech (Shanghai, China) at a purity of 98%. It is stocked in dimethylsulfoxide (DMSO) solution and stored at −30°C. Hyperhomocysteinemia was purchased from Sigma (St. Louis, MO, USA). TRizol was obtained from Invitrogen (California, USA), the LOX‐1, SIRT1 and Caspase‐3 antibody were purchased from Santa Cruz Biotechnology Inc. (California, USA).
Hcy(D090821), DCFH‐DA(D6883), DMSO(D5879), Hepes, SOD, MDA and NADPH oxidase activity kits were got from Sigma‐Aldrich (USA). IL‐6, IL ‐8, CXCL15 and TNF‐α ELISA test kit were purchased from Shanghai Hong LiBiotechnology Company.

Doxorubicin was purchased from Cell Signaling Technology (Danvers, MA, USA). LC3B antibody was purchased from Sigma-Aldrich (St. Louis, MO, USA); caspase-3 antibodies were purchased from Santa Cruz (Dallas, TX, USA) and Cell Signaling Technology (Danvers, MA, USA); p62 antibody and horseradish peroxidase- (HRP-) conjugated secondary antibody were purchased from Cell Signaling Technology (Danvers, MA, USA); CyclinG1 antibody was purchased from Proteintech Group (Chicago, IL, USA). Unless otherwise specified, all other chemicals were purchased from Sigma (St. Louis, MO, USA) or Solarbio (Beijing, China).