The murine macrophage-like cell line RAW 264.7 was obtained from the ATCC. The cells were cultured in Dulbecco’s modified Eagle’s medium (Gibco) supplemented with 10% (v/v) fetal bovine plasma (Gibco) with 100 U/ml penicillin, 100 μg/ml streptomycin (Gibco) and 100 μg/ml Primocin (In vivoGen) at 37°C in a humid atmosphere with 5% CO2. Then, RAW 264.7 cells were reseeded in 96-well or 6-well culture plates at densities of 1×105 or 1×106 cells/well and used for follow-up experiments. RAW 264.7 cells were stimulated with LPS (1 μg/ml) to establish a cell inflammation model and LPS (1 μg/ml) plus zVAD (20 μM; APExBIO) to establish a necroptosis model, with or without KW2449 (1 μM) or RIPA56 (1 μM; MedChemExpress LLC). The cell supernatant was collected 24 h after drug addition, and the cells were collected at the corresponding time points for detection.
Ripa 56
Manufactured by MedChemExpress
Sourced in United States
RIPA-56 is a laboratory equipment product designed for protein extraction and purification. It is a buffer solution that helps to lyse cells and solubilize proteins, allowing for their isolation and analysis. The core function of RIPA-56 is to facilitate the extraction and preparation of protein samples for various downstream applications, such as Western blotting, immunoprecipitation, and other protein analysis techniques.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine plasma, by Thermo Fisher Scientific (1 mentions)
Primocin, by InvivoGen (1 mentions)
Kw2449, by MedChemExpress (1 mentions)
Lipopolysaccharides (lps), by Apexbio (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Necrostatin 1, by MedChemExpress (1 mentions)
Byt c0222, by Beyotime (1 mentions)
Necrosulfonamide, by MedChemExpress (1 mentions)
Hy 100573, by MedChemExpress (1 mentions)
Cisplatin, by MedChemExpress (1 mentions)
Rpmi 1640 medium, by Procell (1 mentions)
3 protocols using ripa 56
1
Murine Macrophage-Like Cell Inflammation and Necroptosis
2
RIPA-56 Intragastric Administration
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RIPA-56 (No. HY-101032; MedChemExpress) was prepared fresh before every use. In total, 2 mg RIPA-56 was placed into a 1.5 mL centrifuge tube, 120 mL dimethyl sulfoxide (DMSO) was immediately added, and the mixture was shaken rigorously. The dissolved RIPA-56 was diluted with 1.08 mL normal saline to make the final working solution. Finally, this diluted RIPA-56 was administered intragastrically at 60 mg kg -1 body weight every day [21] .
3
Oesophageal Cell Lines for Necroptosis
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Oesophageal epithelial cells (HET-1A) (RRID: CVCL_3702) and ESCC cells (KYSE410, KYSE30, KYSE510, KYSE450, and KYSE150) (RRID: CVCL_1352, RRID: CVCL_1351, RRID: CVCL_1354, RRID: CVCL_1353 and RRID: CVCL_1348) were obtained from the Cell Bank of Chinese Academy of Sciences (Shanghai, China) and Procell Life Science&Technology Co., Ltd (Wuhan, China). Cell lines were cultured in RPMI 1640 medium (PM150110, Procell, China) with 10% fetal bovine serum (164210, Procell, Wuhan, China), penicillin G (100 U/ml, BYT-C0222, Beyotime, China), and streptomycin (100 μg/ml, BYT-C0222, Beyotime, China) and maintained in a humidified incubator with 5% CO2, at 37 °C. All cell lines tested negative for mycoplasma throughout the study. The short tandem repeat (STR) profiling was used to verify the identity of each cell line. Cisplatin (HY-17394, MedChemExpress, Monmouth Junction, NJ, USA), necrostatin-1(Nec-1) (HY-15760, MedChemExpress, Monmouth Junction, NJ, USA), RIPA-56 (HY-101032, MedChemExpress, Monmouth Junction, NJ, USA) and necrosulfonamide (NSA) (HY-100573, MedChemExpress, Monmouth Junction, NJ, USA) were used to treat cells for 24 h to induce and inhibit necroptosis, respectively.
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