U251 and DBTRG cells were seeded into 96-well plates (2 × 104 cells/well) and cultured for 12 h. After washing, U251 and DBTRG cells were incubated with 10% CCK-8 (Dojindo Molecular Technologies, Inc., Minato-ku, Tokyo, Japan) and optical density measured using a xMark Microporous Plate Absorption Spectrophotometer (Bio-Rad Laboratories, Inc., Hercules, CA, USA).
Xmark microporous plate absorption spectrophotometer
Manufactured by Bio-Rad
Sourced in Japan
The XMark Microporous Plate Absorption Spectrophotometer is a laboratory instrument designed for quantitative analysis of samples. It measures the absorption of light by samples in a microplate format, enabling multiple samples to be analyzed simultaneously.
Automatically generated - may contain errors
Lab products found in correlation
Cck 8, by Dojindo Laboratories (3 mentions)
As2o3, by Merck Group (1 mentions)
Novocyte 1040 flow cytometer, by Agilent Technologies (1 mentions)
Total glutathione peroxidase assay kit, by Beyotime (1 mentions)
Total superoxide dismutase assay kit, by Beyotime (1 mentions)
Lipid peroxidation mda assay kit, by Beyotime (1 mentions)
Reactive oxygen species assay kit, by Solarbio (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
5 protocols using xmark microporous plate absorption spectrophotometer
1
Cell Viability Assay for U251 and DBTRG Cells
2
Arsenic trioxide induces apoptosis in T24 cells
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T24 cells were seeded into 96-well plates (2 × 104 cells/well) and cultured for 12 h. Next, T24 cells were incubated with As2O3 (Sigma-Aldrich, Merck KGaA, Darmstadt, Germany) at 0, 10, and 20 μmol/L for 6 h. After washing, T24 cells were incubated with 10% CCK-8 (Dojindo Molecular Technologies, Inc., Minato-ku, Tokyo, Japan) and optical density measured using a xMark Microporous Plate Absorption Spectrophotometer (Bio-Rad Laboratories, Inc., Hercules, CA, USA).
The apoptotic rate of T24 cells was detected using an Annexin V, 633 Apoptosis Detection Kit (Dojindo Molecular Technologies), following the kit instructions. T24 cells were seeded into 6-well plates (5 × 105 cells/well) and cultured for 12 h. Next, cells were incubated with As2O3 (Sigma-Aldrich) at 20 μmol/L for 6 h, then incubated with Annexin V, followed by propidium iodide (PI) buffer for 15 min at 25°C in a dark room. Subsequently, apoptotic cells were quantified using a NovoCyte 1040 flow cytometer (ACEA Biosciences, Inc., Zhejiang, China).
The apoptotic rate of T24 cells was detected using an Annexin V, 633 Apoptosis Detection Kit (Dojindo Molecular Technologies), following the kit instructions. T24 cells were seeded into 6-well plates (5 × 105 cells/well) and cultured for 12 h. Next, cells were incubated with As2O3 (Sigma-Aldrich) at 20 μmol/L for 6 h, then incubated with Annexin V, followed by propidium iodide (PI) buffer for 15 min at 25°C in a dark room. Subsequently, apoptotic cells were quantified using a NovoCyte 1040 flow cytometer (ACEA Biosciences, Inc., Zhejiang, China).
3
Cell Proliferation Assay with CCK-8
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Fadu and Cal27 cells were seeded into 96-well plates (2 × 104 cells/well) and cultured for 12 h. After washing, the Fadu and Cal27 cells were incubated with 10% CCK-8 (Dojindo Molecular Technologies, Inc., Minato-ku, Tokyo, Japan), and the optical density was measured using a xMark Microporous Plate Absorption Spectrophotometer (Bio–Rad Laboratories, Inc., Hercules, CA, USA).
4
Oxidative Stress Measurement in T24 Cells
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Total ROS levels in T24 cells were measured using a reactive oxygen species assay kit (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China). T24 cells were collected and suspended in diluted 2,7-dichlorofluorescin diacetate (1 × 107 cells/mL). After incubation for 20 min and washing with culture medium (serum-free), fluorescence intensity was detected at 525 nm emission and 490 nm excitation. Levels of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) in T24 cell lysates were measured using standard assay kits, including the Lipid Peroxidation MDA Assay Kit (Beyotime), the Total Superoxide Dismutase Assay Kit (Beyotime), and the Total Glutathione Peroxidase Assay Kit (Beyotime), according to their respective instructions. Enzyme levels and activity were measured using an xMark Microporous Plate Absorption Spectrophotometer (Bio-Rad).
5
Investigating RNF185 and miR-587 in Glioblastoma
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Glioblastoma cells lines U87, U251 and DBTRG were purchased from the Cell Bank Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). Cells were cultured in DMEM (Gibco, Carlsbad, CA, USA) supplemented with 10% of fetal bovine serum (Gibco) and 1% of penicillinstreptomycin at 37°C, 5% CO 2 humidi ed atmosphere. The full-length sequence of RNF185 mRNA was cloned into pcDNA3.1 vector, and transfected into glioblastoma cells, according to manufacturer's instructions. MiR-587 inhibitors were added into glioblastoma cells. The overexpression and knockdown e ciency were validated with qRT-PCR. GAPDH and U6 were used as the house keeping gene. Primers for RNF185, miR-587, GAPDH and U6 are as follows: RNF185: F-5'-GTGTTTACATCAGTGGTTGGAGA-3'; R-5'-GTGCTGCCCCTTCCATAGAG-3'; GAPDH: F-5'-GGAGCGAGATCCCTCCAAAAT-3'; R-5'-GGCTGTTGTCATACTTCTCATGG - 3'; miR-587: F-5'-CCAGGCAAGAGAGAGTTGCTG-3'; R-5'-AGTCACAGGTGCAGACACATT-3'; U6: F-5'-CTCGCTTCGGCAGCACA'; R-5'-AACGCTTCACGAATTTGCGT-3'.
Cell Counting Kit-8 (CCK-8) U251 and DBTRG cells were seeded into 96-well plates (2×10 4 cells/well) and cultured for 12 h. After washing, U251 and DBTRG cells were incubated with 10% CCK-8 (Dojindo Molecular Technologies, Inc., Minato-ku, Tokyo, Japan) and optical density measured using a xMark Microporous Plate Absorption Spectrophotometer (Bio-Rad Laboratories, Inc., Hercules, CA, USA).
Cell Counting Kit-8 (CCK-8) U251 and DBTRG cells were seeded into 96-well plates (2×10 4 cells/well) and cultured for 12 h. After washing, U251 and DBTRG cells were incubated with 10% CCK-8 (Dojindo Molecular Technologies, Inc., Minato-ku, Tokyo, Japan) and optical density measured using a xMark Microporous Plate Absorption Spectrophotometer (Bio-Rad Laboratories, Inc., Hercules, CA, USA).
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