材料与试剂
- 15 ml离心管(Corning, catalog number: CLS430791)
- 50 ml离心管(Corning, catalog number: CLS430828)
- 10 ml移液管(Corning, catalog number: 4488)
- 1.5 ml离心管(Axygen, catalog number: MCT-150-C)
- 96孔圆底板(Corning, catalog number: CLS3879)
- 384孔黑色板(Corning, catalog number: COR-3712)
- RPMI 1640(Thermo, catalog number: 11875093)
- Opti-MEM(Thermo, catalog number: 31985088)
- FBS(Biological Industries, catalog number: 04-001-1A)
- Trypsin-EDTA(0.25%)(Thermo, catalog number: 25200072)
- Penicillin/streptomycin(Thermo, catalog number: 15070063)
- Lipofectamine RNAiMAX reagent(Thermo, catalog number: 13778075)
- RSL3 reagent(MCE, catalog number: HY-100218A)
- Ferrostatin-1 reagent(MCE, catalog number: HY-100579)
- DMSO reagent(Sigma, catalog number: D2650)
- Propidium Iodide reagent(Thermo, catalog number: P3566)
- Hoechst 33342 reagent(Thermo, catalog number: P3570)
- 1640完全细胞培养液(见溶液配方)
- PBS缓冲液(见溶液配方)
细胞系
HGC-27细胞(上海生命科学研究院细胞资源中心,TCHu 22)
仪器设备
- Multidrop自动分液器(Thermo Fisher,model: Multidrop Combi)
- 隔水式恒温培养箱(上海精宏实验设备有限公司)
- 水平冷冻离心机(Eppendorf,model: 5810R)
- 细胞计数仪(Invitrogen,model: Countess)
- 二氧化碳恒温细胞培养箱(Thermo Fisher,model: Thermo Series 8000)
- 电动移液器(Eppendorf)
- Ensight多功能微孔板检测仪(Perkin Elmer,model: Ensight)
- 非接触式数码滴定仪(Tecan,model: D300e)
- Beckman大型液体工作站(Beckman,model: Biomek FXP)
实验步骤