Ldh cytotoxicity kit

Manufactured by Beyotime

Sourced in China

The LDH cytotoxicity kit is a laboratory tool used to measure the release of lactate dehydrogenase (LDH) from damaged or lysed cells. It provides a quantitative assessment of cytotoxicity or cell viability in various in vitro experimental settings.

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Lab products found in correlation

Microplate reader, by Agilent Technologies (2 mentions) Cell counting kit 8 cck 8, by Beyotime (2 mentions) Microplate reader, by Thermo Fisher Scientific (2 mentions) Model 550 microplate reader, by Bio-Rad (2 mentions) Cck 8 assay kit, by Apexbio (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Wuhan Servicebio Technology (1 mentions) Fluorescence microscope, by Leica (1 mentions) Fluo 4 am, by Thermo Fisher Scientific (1 mentions) Gsh px assay kit, by Solarbio (1 mentions) Sod assay kit, by Solarbio (1 mentions) Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions) Triton x 100, by Merck Group (1 mentions) Imark microplate reader, by Bio-Rad (1 mentions)

Close spelling variants of this product

Lactate dehydrogenase (LDH) cytotoxicity assay kit LDH release quantification cytotoxicity Assay Kit LDH cytotoxicity assay detection kit LDH cytotoxicity test kit LDH cytotoxicity detection kit Lactate dehydrogenase (LDH) cytotoxicity detection kit LDH Cytotoxicity Assay Kit (C0016) LDH Cytotoxicity Assay Kit LDH Cytotoxicity Assay Kit II Lactate Dehydragenase (LDH) Cytotoxicity Assay Kit

13 protocols using ldh cytotoxicity kit

Cytoprotective Effect of Tanshinone IIA on DON-Induced Toxicity

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Cell viability was measured by a CCK8 assay kit (APExBIO, Houston, TX, USA). IPEC-J2 cells were seeded at a density of 2 × 10⁵ cells/well in a 96-well plate. First, the cells were incubated with different concentrations of Tan IIA (0, 15, 30, 45, 60 and 75 μg/mL) to select the optimal concentration of Tan IIA. Then, the cells were incubated with 1 μM DON and/or optimal Tan IIA to explore the protective effect of Tan IIA on DON-induced IPEC-J2 cell damage. Finally, the cell viability value was analysed by a microplate reader (TECAN, Männedorf, Switzerland).
Lactate dehydrogenase (LDH) release was measured by an LDH cytotoxicity kit (Beyotime, Shanghai, China). After incubation with 1 μM DON and/or 45 μg/mL Tan IIA, the supernatants were collected and treated with LDH reagents. Finally, LDH release was analysed by a microplate reader (TECAN, Switzerland).

Zhang C., Wang Y., Zhang X., Zhang K., Chen F., Fan J., Wang X, & Yang X. (2024). Maintaining the Mitochondrial Quality Control System Was a Key Event of Tanshinone IIA against Deoxynivalenol-Induced Intestinal Toxicity. Antioxidants, 13(1), 121.

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Assessing AgNPs Cytotoxicity on PMNs

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PMNs (5 ×10⁵ cells per well) were incubated with various concentrations (0.5 μg/mL, 1 μg/mL, 2 μg/mL, and 4 μg/mL) of AgNPs in complete RPMI 1640 medium (HyClone, USA) for 4 h. Then, the co-culture medium of each well was replaced with 0.1 mL of serum-free RPMI 1640 medium containing 10 μL of CCK-8 solution (Dojindo Kagaku Co, Kumamoto, Japan), followed by incubation for 1 h. Next, the absorbance of the incubated solution system was determined at 450 nm using a microplate reader (Thermo Scientific, USA). Furthermore, lactate dehydrogenase (LDH) assays were used to determine the cellular activity of PMNs using the LDH cytotoxicity kit (Beyotime, Shanghai, China) according to the manufacturer’s protocol. In addition, after coculture with AgNPs for 4 h, PMNs were fixed with 4% paraformaldehyde for 10 min and permeabilized with 0.1% Triton-X for 5 min, and then stained with TRITC-phalloidin and Dapi (Servicebio, Wuhan, China) for 30 and 10 min, respectively. Fluorescent images were obtained using a fluorescence microscope (Leica, Hamburg, Germany).

Huang M., Ye K., Hu T., Liu K., You M., Wang L, & Qin H. (2021). Silver Nanoparticles Attenuate the Antimicrobial Activity of the Innate Immune System by Inhibiting Neutrophil-Mediated Phagocytosis and Reactive Oxygen Species Production. International Journal of Nanomedicine, 16, 1345-1360.

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Astrocyte Viability and Cytotoxicity Assay

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Cell viability and LDH was determined with Cell Counting Kit-8 (CCK-8, Beyotime, China) and LDH cytotoxicity kit (Beyotime) under the manufacturer's instructions. Astrocytes were plated into 96/24-well plate, with or without BM-MSCs coculture, treated with hemin for another 24 h. The absorbance at 450 nm and 490 nm was read with the microplate reader (BioTek, USA).

Chen X., Xu C.X., Liang H., Xi Z., Pan J., Yang Y., Sun Q., Yang G., Sun Y, & Bian L. (2020). Bone marrow mesenchymal stem cells transplantation alleviates brain injury after intracerebral hemorrhage in mice through the Hippo signaling pathway. Aging (Albany NY), 12(7), 6306-6323.

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Cytotoxicity and Intracellular Ca2+ Assay

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Lactate dehydrogenase (LDH) activity was measured to estimate the cytotoxicity with a LDH Cytotoxicity Kit (Beyotime, China). The level of intracellular Ca2+ was determined by flow cytometry after being incubated with fluorescent Fluo-4/AM according to the recommendations from the manufacturer (Invitrogen, United States).

Mei M., Sun H., Xu J., Li Y., Chen G., Yu Q., Deng C., Zhu W, & Song J. (2022). Vanillic acid attenuates H2O2-induced injury in H9c2 cells by regulating mitophagy via the PINK1/Parkin/Mfn2 signaling pathway. Frontiers in Pharmacology, 13, 976156.

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Quantifying Cellular Stress Responses

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Following treatment as described above, the cells were collected and centrifuged at 3,000 × g for 10 min at 4°C, and the supernatant was stored at −80°C. The LDH release levels, MDA levels and SOD and GSH-Px activity were detected using a LDH cytotoxicity kit (Beyotime Institute of Biotechnology), and MDA assay kit (Nanjing Jiancheng Institute of Bioengineering Institute), an SOD assay kit (Beijing Solarbio Science & Technology Co., Ltd.) and a GSH-Px assay kit (Beijing Solarbio Science & Technology Co., Ltd.), respectively. All assays were performed strictly according to the manufacturer's protocol.

Liu B., Wei H., Lan M., Jia N., Liu J, & Zhang M. (2020). MicroRNA-21 mediates the protective effects of salidroside against hypoxia/reoxygenation-induced myocardial oxidative stress and inflammatory response. Experimental and Therapeutic Medicine, 19(3), 1655-1664.

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NK Cell-Mediated Cytotoxicity Assay

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This assay was performed using an LDH cytotoxicity kit (C0016, Beyotime). GSCs (target cells, 5 × 10⁴ cells/ml) were cocultured for 2 h with NK-92MI or primary NK cells (effector cells) at T:E ratios of 1:2, 1:5 and 1:10 in 96-well plates. NKG2D blocking antibody (BE0351, BioXcell) or isotype IgG control was added at a concentration of 10 μg/ml for specific experiments. After incubation, supernatants from each well were collected for further analysis and calculation according to the manufacturer’s instructions.

Zhong J., Yang X., Chen J., He K., Gao X., Wu X., Zhang M., Zhou H., Xiao F., An L., Wang X., Shi Y, & Zhang N. (2022). Circular EZH2-encoded EZH2-92aa mediates immune evasion in glioblastoma via inhibition of surface NKG2D ligands. Nature Communications, 13, 4795.

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Evaluating Astrocyte Viability with CCK-8

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Cell viability or LDH was evaluated with Cell Counting Kit-8 (CCK-8, Beyotime, China) or LDH cytotoxicity kit (Beyotime) following the manufacturer’s instructions. Astrocytes were plated into 96/24-well plate, with or without BM-MSCs coculture. The cells were treated with hemin for another 24 h. Then the experiment continued according to the instructions of the kit. The absorbance at 450 nm/490 nm was read with a microplate reader (BioTek, United States). The result was expressed as the percentage of the control group.

Chen X., Liang H., Xi Z., Yang Y., Shan H., Wang B., Zhong Z., Xu C., Yang G.Y., Sun Q., Sun Y, & Bian L. (2020). BM-MSC Transplantation Alleviates Intracerebral Hemorrhage-Induced Brain Injury, Promotes Astrocytes Vimentin Expression, and Enhances Astrocytes Antioxidation via the Cx43/Nrf2/HO-1 Axis. Frontiers in Cell and Developmental Biology, 8, 302.

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AgNPs Modulate Neutrophil Cytotoxicity

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Isolated PMNs were cocultured with different dosages (0.5 μg/mL, 1 μg/mL, and 2 μg/mL) of AgNPs and 1×10⁷ CFU/mL bacteria. After incubation for 2 h, the PMN lysis rate of each group was measured using the LDH cytotoxicity kit (Beyotime, Shanghai, China) as mentioned above.

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H2O2-Induced Cell Viability Assay

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After being stimulated by 600 μM of H₂O₂, cell viability inhibition was determined by the MTT (methylthiazolyldiphenyl-tetrazolium bromide, MTT, Sigma (United States) assay. MTT was dissolved in 0.8% NaCl solution to a concentration of 5 mg/ml. Lactate dehydrogenase (LDH) activity was measured to estimate the cytotoxicity with an LDH Cytotoxicity Kit (Beyotime, China).

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Lactate Dehydrogenase Cytotoxicity Assay

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The cell treatment was as described for the MTT assay (above). Cell culture medium was used as a blank control. Untreated MCF-7 cells were used as a spontaneous release control. Normal MCF-7 cells treated with Triton X-100 (Sigma-Aldrich, St. Louis, USA) were used as a maximum release control. In accordance with the instructions of the lactate dehydrogenase (LDH) cytotoxicity kit (Beyotime Biotechnology, Shanghai, China), 120 μL of supernatant from each well was collected and transferred to a new 96-well plate. Then 60 μL of LDH working reaction mixture was added to each well and incubated in the dark at room temperature for 30 min. The absorbance of each well was measured at a wavelength of 490 nm. Each sample had 3 replicate wells. For each test sample the cytotoxicity (%) = (experimental value -spontaneous release)/(maximum release -spontaneous release) × 100%.

Li G., Xu D., Sun J., Zhao S, & Zheng D. (2020). Paclitaxel inhibits proliferation and invasion and promotes apoptosis of breast cancer cells by blocking activation of the PI3K/AKT signaling pathway. Advances in clinical and experimental medicine : official organ Wroclaw Medical University, 29(11).

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