The tumor cell line Yac-1 was obtained from the American Type Culture Collection (Rockville, MD). Splenocytes were collected from mice in animal experiment 1 and cocultured with Yac-1 cells to obtain an E : T (splenocytes : Yac-1) ratio of 75 : 1 in V-bottomed 96-well plates. After 4 h, cytotoxicity was determined by the lactate dehydrogenase (LDH) assay using a LDH Cytotoxicity Assay Kit (Roche, Basel, Switzerland).
Ldh cytotoxicity assay kit
Manufactured by Roche
Sourced in United States, Germany, Switzerland
The LDH Cytotoxicity Assay Kit is a quantitative colorimetric assay that measures lactate dehydrogenase (LDH) activity released from damaged cells. It provides a simple, accurate, and sensitive method for determining cytotoxicity or cell death in cell culture systems.
Automatically generated - may contain errors
Lab products found in correlation
In situ cell death detection kit, by Roche (3 mentions)
Jc 1 mitochondrial membrane potential assay kit, by Cayman Chemical (2 mentions)
Yac 1, by American Type Culture Collection (1 mentions)
Alkaline phosphatase assay kit, by Nanjing Jiancheng (1 mentions)
Cck 8 kit, by Solarbio (1 mentions)
Resazurin salt, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Annexin v 7 amino actinomycin d 7 aad apoptosis detection kit, by BD (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Amphotericin b, by Thermo Fisher Scientific (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
Low gelling temperature agarose, by Merck Group (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Dulbecco s minimal essential medium dmem, by Thermo Fisher Scientific (1 mentions)
Protease inhibitor cocktail p1860, by Merck Group (1 mentions)
Earle s balanced salt solution ebss, by Merck Group (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Triton x 100, by Merck Group (1 mentions)
31 protocols using ldh cytotoxicity assay kit
1
Cytotoxicity Assay of Yac-1 Cells
2
Evaluating Syringaldehyde Cytotoxicity in HeLa Cells
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HeLa cells were plated into 96‐well plate at a density of 2×104 cells/well and were incubated at 37°C in a 5% CO2 incubator for 16‐18 hours before infection. After washing the cells three times with PBS, they were infected at an MOI of 100 with S. Typhimurium and cultured with different concentrations of syringaldehyde for 5 hours in DMEM. Subsequently, the cultures were centrifuged at 1000 rpm for 10 min and the LDH release was then measured in culture supernatants on a microplate reader at 490 nm using an LDH Cytotoxicity Assay Kit according to the manufacturer's instructions (Roche, Switzerland). The percentage of LDH release was calculated as described in a previous study.17 The LDH release from the HeLa cells incubated with syringaldehyde was measured at an effective concentration range (0.0225‐0.18 mmol/L) in independent tests to evaluate the cytotoxicity of syringaldehyde.
3
NK Cell-Mediated Cytotoxicity Assay
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Activated spleen NK cells were isolated and co-cultured with Yac-1 cells from a mouse lymphoma cell line in V-bottomed 96-well plates in 200 μL of RPMI 1640 10% fetal calf serum for 24 h. 50,000 target Yac-1 cells were co-cultured in triplicate with 5000 (1:10), 25,000 (1:2), 50,000 (1:1), 250,000 (5:1), and 500,000 (10:1) NK cells, respectively. Cytotoxicity was determined by the lactate dehydrogenase (LDH) assay. The LDH assay was performed using a LDH Cytotoxicity Assay Kit (Roche, Basel, Switzerland) according to the manufacturer’s instructions.
4
Quantifying Lung Enzyme Levels
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The amounts of alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) released in the BALF were determined by Alkaline Phosphatase Assay Kit (Nanjing Jiancheng) and LDH Cytotoxicity Assay Kit (Hoffman-La Roche Ltd., Basel, Switzerland) according to the manufacturers’ instructions.
5
Cytotoxicity and Cell Viability Assays
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Cell lactate dehydrogenase (LDH) content was measured with a LDH Cytotoxicity Assay Kit (Roche) as described.33 Cell viability was measured using the CCK‐8 kit (Solarbio). In brief, H9C2 cells were seeded at 5 × 103 cells per well in 96‐well plates in triplicate. After the transfection and 48 hours treatments, 10 μL of CCK‐8 solution was added to each well. After 2 hours of incubation, absorbance was measured at 450 nm.
6
LDH Release Assay for Bacterial Cell Membrane Activity
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The LDH release
assay was performed to verify the cell membrane activity of bacterial
cells treated with BQAS using the LDH cytotoxicity assay kit (Roche
Applied Science).47 (link) The standard protocol
assay was conducted according to the manufacturer’s instructions.
Briefly, the E. coli and S. aureus cells were treated with BQAS at concentrations
of 12.5, 25, 50, 100, and 200 μg/mL for 3 h, followed by transferring
120 μL of each cell culture supernatant into a new centrifuge
tube and then adding 60 μL of substrate. After incubation at
room temperature in the dark for 30 min, the reaction was stopped
by adding 50 μL of the stop solution. Finally, the LDH release
was quantified by monitoring the absorbance at 490 nm.
assay was performed to verify the cell membrane activity of bacterial
cells treated with BQAS using the LDH cytotoxicity assay kit (Roche
Applied Science).47 (link) The standard protocol
assay was conducted according to the manufacturer’s instructions.
Briefly, the E. coli and S. aureus cells were treated with BQAS at concentrations
of 12.5, 25, 50, 100, and 200 μg/mL for 3 h, followed by transferring
120 μL of each cell culture supernatant into a new centrifuge
tube and then adding 60 μL of substrate. After incubation at
room temperature in the dark for 30 min, the reaction was stopped
by adding 50 μL of the stop solution. Finally, the LDH release
was quantified by monitoring the absorbance at 490 nm.
7
Quantifying NET Reduction by ESP Cytotoxicity
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Lactate dehydrogenase (LDH) released from PMA-treated PMNs was quantified to determine whether NET reduction was due to ESP cytotoxicity. Briefly, PMNs were inoculated into a 96-well cell culture plate at 1 × 105 cells/well, and different concentrations of Ad3 ESP (2, 5, and 10 ng/μl) with 25 μM ATA were added into the culture medium. After 1 h incubation, PMNs were stimulated with 100 nM PMA for 3 h. All steps were maintained at 37 °C with 5% CO2. The plate was centrifuged at 400g for 5 min, and 120 μl supernatant from each well was transferred to a new 96-well plate. The LDH activity in the supernatants was determined using an LDH Cytotoxicity Assay Kit (Roche Applied Science, Germany) according to the manufacturer's protocol after incubation in darkness for 30 min. Samples included the experimental group, a background control, negative control of unstimulated PMN (0% cell lysis), and positive control of maximum LDH release (100% cell lysis).
8
Measuring BMDM Cytotoxicity by LDH
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BMDMs were incubated for 3 h with purified SAK or a 1/20 or 1/100 dilution of each S. aureus supernatant (stationary phase). BMDMs lysis was measured using a lactate dehydrogenase (LDH) cytotoxicity assay kit according to the manufacturer’s protocol (Roche). The released LDH was normalized to the total LDH content measured in BMDMs samples infiltrated with 2% Triton X-100.
9
Cell Viability and Cytotoxicity Assay
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RPMI 1640 medium and fetal bovine serum were from Gibco. Amphotericin B was from Invitrogen Life Technologies. Resazurin salt, propidium iodide, and RNase were from Sigma Aldrich. Annexin V/7-amino-actinomycin D (7-AAD) apoptosis detection kit was obtained from BD Bioscience. LDH cytotoxicity assay kit was from Roche. Fast staining kit (RAL-555) was from RAL Reactifs. All other reagents are of analytical grade from different commercial sources.
10
Quantifying Oxidative Stress-Induced Cardiac Injury
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To detect ROS-induced cell injury, the treated H9c2 were co-cultured with Dichloro-dihydro-fluorescein diacetate (DCFH-DA in the dark for 40 min and then observed by scanning confocal microscopy. The creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH) were detected via the CK-MB isoenzyme assay kit and LDH Cytotoxicity Assay Kit (Roche, United States), respectively. Besides, the concentrations of interleukin (IL)-6 and TNF-ɑ were assessed using ELISA kits (Roche, Basel, Switzerland).
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