Goat igg hrp secondary antibodies against rabbit and mouse

Cells or tumor tissues were suspended in Laemmli Buffer (Bio-Rad Laboratory, USA). Protein concentrations were measured and normalized using the BCA Protein Assay Kit (Pierce, USA). Equal amounts of protein samples were electrophoresed by SDS-PAGE and were transferred to polyvinylidene difluoride transfer membranes (Bio-Rad Laboratory, USA). The membrane was blocked with 5% fresh nonfat milk in TBST and was incubated with specific primary antibodies in TBST overnight at 4°C. The membrane was washed 3 times with TBST and incubated with secondary antibodies for 2 hours at room temperature. Finally, the blot bands were visualized using an ECL kit (Bio-Rad, Hercules, CA). The Bcl-2, Bcl-xL, BAX, Caspase 3 and 9 antibodies were from Cell Signaling Technology (MA, USA), and the GAPDH, c-Myc, Cyclin D and Cyclin E antibodies were from Abcam (Cambridge, UK); the goat IgG-HRP secondary antibodies against rabbit and mouse were from Zhongshan Golden Bridge Biotechnology (Beijing, China).

SHK was purchased from Sigma-Aldrich (St. Louis, MO, USA), dissolved in DMSO (Sigma-Aldrich, St. Louis, MO, USA). ZVAD-FMK, Necrostatin-1, 5-Fluorouracil and Oxaliplatin were from MedChemExpress (NJ, USA). ZDEVD-FMK and ZLEHD-FMK were from Adooq bioscience (CA, USA). N-acetyl-L-cysteine and L-glutathione were from Beyotime Institute (Shanghai, China). Antibodies were as follows: PARP, Caspase 3, Caspase 8, Caspase 9, RIPK1, Cytochrome C and VDAC1 were from Cell Signaling Technology (MA, USA); AIF, Endonuclease G, GAPDH and PCNA were from Abcam (Cambridge, UK); the goat IgG-HRP secondary antibodies against rabbit and mouse were from Zhongshan Golden Bridge Biotechnology (Peking, China) and FITC or Cy3 conjugated secondary antibodies were from Sigma (MO, USA).