A2494001

Manufactured by Merck Group

Sourced in United States

The A2494001 is a laboratory equipment product. It is designed to perform a specific core function, but a detailed description is not available while maintaining an unbiased and factual approach without extrapolation.

Automatically generated - may contain errors

Lab products found in correlation

Hcc1806, by Thermo Fisher Scientific (1 mentions) S11550, by R&D Systems (1 mentions) Sum159pt, by BioIVT (1 mentions) Sum159pt, by Thermo Fisher Scientific (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Cc 3150, by Lonza (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Sartorius (1 mentions) Hbzy 1, by National Collection of Authenticated Cell Cultures (1 mentions) Resazurin, by Merck Group (1 mentions) G7513, by Merck Group (1 mentions) Enzychrom nad nadh assay kit, by BioAssay Systems (1 mentions) A1443001, by Thermo Fisher Scientific (1 mentions)

3 protocols using a2494001

Comprehensive Breast Cancer Cell Culture

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All cell lines were purchased directly from ATCC (American Type Culture Collection) except for SUM159pt which was purchased from BioIVT. MDA-MB-231, MDA-MB-468, HCC1806 and T47D cells were cultured in RPMI-1640 (Gibco, 11879020) supplemented with 5 mM glucose (Gibco, A2494001), 10% fetal bovine serum (R&D, S11550), and 1x antibiotic-antimycotic solution (Corning, 30-004-CI). SUM159pt cells were cultured in RPMI-1640 (Gibco, 11879020) supplemented with 5 mM glucose (Gibco, A2494001), 5ug/ml Insulin (Sigma-Aldrich, I9278), 1ug/ml hydrocortisone (Sigma-Aldrich, H4001), 5% fetal bovine serum (R&D, S11550), and 1x antibiotic-antimycotic solution (Corning, 30-004-CI). MCF10A cells were cultured in MEGM (Lonza, CC-3150) supplemented with 1x antibiotic-antimycotic solution (Corning, 30-004-CI). All the cells were incubated at 37°C with 5% CO₂ and 95% relative humidity.

Baek M.L., Lee J., Pendleton K.E., Berner M.J., Goff E.B., Tan L., Martinez S.A., Mahmud I., Wang T., Meyer M.D., Lim B., Barrish J.P., Porter W., Lorenzi P.L, & Echeverria G.V. (2023). Mitochondrial structure and function adaptation in residual triple negative breast cancer cells surviving chemotherapy treatment. Oncogene, 42(14), 1117-1131.

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Mesangial Cell Viability under Glucose and MCC950

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The rat mesangial cell line HBZY-1 was purchased from the China Center for Type Culture Collection (Wuhan, China) and were cultured in low-glucose (5.5 mmol/L) DMEM (Hyclone, SH30021, USA) with 10% FBS (Bioind, 04–001-1A, USA), 100 μg/mL streptomycin and 100 U/mL penicillin (Gibco, 15140-122, USA). The cells were routinely cultured at 5% CO₂ and 37°C with saturated humidity, digested and passaged after 85% cell confluence. At 24 hrs after cell passaging and attachment, the cells were divided into five groups and cultured for 48 hrs: 1) the normal-glucose group (NG) that received 5.5 mmol/L glucose (Sigma, A24940-01,USA); 2) the high-glucose group (HG) that received 30 mmol/L glucose with vehicle (DMSO); 3) the 0.01 μM MCC950 group (0.01) that received 30 mmol/L glucose with 0.01 μmol/L MCC950; 4) the 0.1 μM MCC950 group (0.1) that received 30 mmol/L glucose with 0.1 μmol/L MCC950 and 5) the 1 μM MCC950 group (1) that received 30 mmol/L glucose with 1 μmol/L MCC950. We tested cell viability at 0 and 48 hrs by light microscopy and trypan blue exclusion.

Zhang C., Zhu X., Li L., Ma T., Shi M., Yang Y, & Fan Q. (2019). A small molecule inhibitor MCC950 ameliorates kidney injury in diabetic nephropathy by inhibiting NLRP3 inflammasome activation. Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy, 12, 1297-1309.

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Quantifying Cellular NAD+ and NADH Levels

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Total cellular NADH and NAD⁺ levels were determined using the EnzyChrom NAD⁺/NADH assay kit (BioAssay Systems, E2ND-100). Briefly, HKC-8 cells were plated at a density of 400,000 cells in a 6-well plate. Next day, cells were washed with PBS and grown in DMEM (ThermoFisher, A1443001) supplemented with 10% FBS, 1x Penicillin/Streptomycin and various carbon sources for 4 h (1) Complete medium contained 5.5 mM glucose (Gibco™, A2494001) and 4 mM glutamine (Sigma-Aldrich, G7513), (2) glutamine free medium contained only 5.5 mM glucose and (3) pyruvate supplemented medium contained, 5.5 mM glucose, 4 mM glutamine and 3 mM sodium pyruvate (Gibco™, 11360070). DNP (25 uM) (Sigma-Aldrich, D198501) and resazurin (10 uM or 50 uM) (Sigma-Aldrich, R7017) treatments were performed for 1 h. Cells were washed with PBS. NAD⁺ and NADH were extracted from the lysate according to the manufacturer’s protocol. The change in absorbance at 565 nm for 15 min at room temperature was measured.

Vellama H., Eskla K.L., Eichelmann H., Hüva A., Tennant D.A., Thakker A., Roberts J., Jagomäe T., Porosk R., Laisk A., Oja V., Rämma H., Volke V., Vasar E, & Luuk H. (2023). VHL-deficiency leads to reductive stress in renal cells. Free Radical Biology & Medicine, 208, 1-12.

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