HepG2 cells (4 × 103) were seeded in a 96-well plate. After an overnight cultivation, cells were treated with different concentrations of curcumin for 6, 12, 24, 48, and 72 h. The non-radioactive cytotoxicity lactate dehydrogenase release assay kit (Promega, USA) was used to measure the cytotoxicity of curcumin against cancer cells in concentrations of 10, 20, 30, 40, 50, 60, and 80 μmol, according to the manufacturer’s protocol. Specific lysis was calculated according to the following formula: percent specific lysis = [(experimental release value − effector spontaneous release value − target spontaneous release value)/(target maximum release value − target spontaneous release value)] × 100%. The results shown are representative of experiments repeated three times.
Lactate dehydrogenase release assay kit
Manufactured by Promega
Sourced in United States
The Lactate dehydrogenase release assay kit is a tool used to measure the activity of the enzyme lactate dehydrogenase (LDH) released from damaged or lysed cells. The kit provides reagents and a protocol to quantify LDH release, which can be an indicator of cytotoxicity or cell membrane integrity.
Automatically generated - may contain errors
Lab products found in correlation
Flow cytometry apparatus, by BD (1 mentions)
Sb203580, by Merck Group (1 mentions)
Cd3 percp, by BD (1 mentions)
Igg1 pe, by BD (1 mentions)
Igg1 fitc, by BD (1 mentions)
Cd3 pe, by BD (1 mentions)
Cd56 fitc, by BD (1 mentions)
Nkg2d pe, by BD (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Ly294002, by Merck Group (1 mentions)
Erlotinib, by Roche (1 mentions)
3 protocols using lactate dehydrogenase release assay kit
1
Curcumin Cytotoxicity in HepG2 Cells
2
Cytotoxicity Assay of NK Cells
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The lactate dehydrogenase-release assay kit (Promega, WI, USA) was used to measure the cytotoxicity of NK cells, according to the manufacturer’s instructions. In brief, NK cells were stimulated with 20 ng/mL of recombinant murine IL-2 for 48 h, washed twice with phosphate-buffered saline, and seeded into 96-well round-bottom microtiter tissue culture plates at various effector:target cell ratios. Target cell samples (1 × 104 cells per well) were tested in triplicate. The cells were incubated for 4 h at 37 °C in a 5% CO2 humidified incubator. Culture supernatants (50 μL) were then collected and combined with 50 μL of the substrate. The plates were covered with aluminum foil for protection against light and incubated at room temperature for 30 min, after which 50 μL of stop solution was added to each well. Absorbance at 490 nm was measured within 1 h of adding the stop solution. The results are expressed as the percentage of specific release based on the following formula: percent specific release = [(experimental release - spontaneous release)/(maximum release - spontaneous release)] × 100.
3
Evaluating Immune Cell Responses in Lung Cancer
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The following materials were used in this study: recombinant human interleukin-2 (Liaoning Weixing Biological Products Institute, Liaoning, China); interferon-g (Shanghai Clone Bio-Tech Co., Ltd., Shanghai, China); lymphocyte separation medium (Tianjin Haoyang Biological Products Co., Ltd., Tianjin, China) ; RPMI 1640 (Gibco, Grand Island, NY, USA); tetrazolium blue (Sigma, St. Louis, MO, USA); CD3 mAb (ProSpec-Tany TechnoGene, Rehovot, Israel); FITC-CD4/PE-CD8/PerCP-CD3, FITC-CD56, PE-CD3, PerCP-CD3, NKG2D-PE, FITC-IgG1, PE-IgG1, MICA, MICB, ULBP1, ULBP2, ULBP3 monoclonal antibodies, and flow cytometry apparatus (BD Biosciences, Franklin Lakes, NJ, USA); lactate dehydrogenase release assay kit (Promega Corporation, Madison, WI, USA); erlotinib (Roche, Basel, Switzerland), LY294002, SB203580 (Sigma), and signal transduction and transcription 21 (STAT21) (Biomol, Farmingdale, NY, USA). The drugs were dissolved in dimethyl sulfoxide and stored at -20°C. Drugs were thawed before use. The RPMI 1640 culture medium containing 10% fetal bovine serum was diluted to the desired concentration. The final concentration of dimethyl sulfoxide was <0.1%. Human lung adenocarcinoma A549 cells were routinely passaged and stored in our center.
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