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Pierce ldh cytotoxicity kit

Manufactured by Thermo Fisher Scientific
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The Pierce™ LDH cytotoxicity kit is a laboratory assay used to quantitatively measure lactate dehydrogenase (LDH) release from damaged cells. It provides a straightforward method for determining cytotoxicity in cell-based assays.

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Lab products found in correlation

Celltiter 96 aqueous one solution cell proliferation assay, by Promega (4 mentions) Varioskan flash reader, by Thermo Fisher Scientific (1 mentions) Mitotracker green fm, by Thermo Fisher Scientific (1 mentions) Urea nitrogen direct kit, by EKF Diagnostics (1 mentions) Ro 32 0432, by Merck Group (1 mentions) Phorbol 12 13 dibutyrate pdbu, by Merck Group (1 mentions) Mito id membrane potential detection kit, by Enzo Life Sciences (1 mentions)

Spelling variants of this product

Pierce LDH Cytotoxicity Assay Kit (430 citations) Pierce Lactate Dehydrogenase (LDH) Cytotoxicity Assay Kit (20 citations) LDH cytotoxicity kit (10 citations) Pierce LDH Cytotoxicity Detection Kit (4 citations) Thermo Scientific Pierce LDH Cytotoxicity Assay Kit (4 citations) LDH kit (3 citations) Pierce lactic dehydrogenase (LDH) cytotoxicity assay kit (2 citations)

10 protocols using pierce ldh cytotoxicity kit

1

Quantifying Cell Death via LDH Release

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Cell death was measured in by release of lactate dehydrogenase (LDH) into the media and normalized to total remaining LDH in the lysate using the Pierce LDH Cytotoxicity Kit (Thermo Fisher Scientific, 88953) according to the manufacturer’s specifications.
Machiela E., Jeloka R., Caron N.S., Mehta S., Schmidt M.E., Baddeley H.J., Tom C.M., Polturi N., Xie Y., Mattis V.B., Hayden M.R, & Southwell A.L. (2020). The Interaction of Aging and Cellular Stress Contributes to Pathogenesis in Mouse and Human Huntington Disease Neurons. Frontiers in Aging Neuroscience, 12, 524369.
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2

Quantifying Cytotoxicity through LDH Release

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The levels of lactate dehydrogenase (LDH) leaked into the cell culture medium were quantified using the Pierce™ LDH cytotoxicity kit (Thermo Fisher) following the manufacturer’s guidelines. In brief, cell supernatant was aspirated after exposure of Calu-3 or TK6 cells to PCP or ZnO ENMs. The aspirated volume was then centrifuged at 3000xg for 30’ to remove any cells, cellular debris, and unbound particles, and the supernatant was mixed with an equal volume of reaction reagents. The reaction was stopped after 30 minutes, and the solution was applied to absorbance measurement at the wavelength of 490 nm (LDH activity) and 680 nm (background signal from instrument). The absorbance value at 680 nm was subtracted from 490 nm prior to calculation. Then, the value from treated samples was subtracted by the data from untreated wells. Relative LDH levels were expressed as the percentage to the LDH levels from cells treated with lysis buffer for maximum amount of released LDH. To assess potential interferences from particles in the Pierce™ LDH assay, culture media alone and particles suspended in culture media at the highest dose were also measured using the same assay.
Bitounis D., Huang Q., Toprani S.M., Setyawati M.I., Oliveira N., Wu Z., Tay C.Y., Ng K.W., Nagel Z.D, & Demokritou P. (2022). Printer center nanoparticles alter the DNA repair capacity of human bronchial airway epithelial cells. NanoImpact, 25, 100379.
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3

Cytotoxicity Assay for Cigarette Smoke Exposure

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Cells were plated at the specified number (10,000, 15,000 or 20,000 cells/well) in 48 well cell culture plates, and grown for 24 hours. Cells were exposed to 400 μL of diluted CSE (0–7%) and were incubated for 24 hours. LDH release was measured using the Pierce LDH cytotoxicity kit (Thermofisher catalog # 88953). Briefly, 50 uL of media was removed from each well, and combined with 50 uL of reaction mixture. This was incubated at room temperature for 30 mins, then 50 uL of stop solution was added. The absorbance was read at 490/680 and LDH release was calculated according to the formula below. Maximum LDH release was obtained by exposing cells to 20% CSE, and spontaneous LDH release was measured in cells exposed to DMEM+10% FBS without CSE:
LDHrelease=100%(CSE−exposedLDHrelease−spontaneousLDHreleaseMaximumLDHrelease−spontaneousLDHrelease)
Bourgeois J.S., Jacob J., Garewal A., Ndahayo R, & Paxson J. (2016). The Bioavailability of Soluble Cigarette Smoke Extract Is Reduced through Interactions with Cells and Affects the Cellular Response to CSE Exposure. PLoS ONE, 11(9), e0163182.
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4

PQ Cytotoxicity on HBMEC

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HBMEC were seeded in a 96-well plate (10,000 cells per well) and treated for 24 h with PQ at different concentrations (0.1, 1, 10, 100, 500 and 1000 µM). Cell proliferation was determined using the MTS assay (CellTiter 96® AQueous One Solution Cell Proliferation Assay, Promega, Madison, WI, USA), whereas cytotoxicity was assessed by measuring lactate dehydrogenase (LDH) released using a Pierce™ LDH cytotoxicity kit (Thermo Scientific, Rockford, IL, USA). Both the MTS and LDH assays were performed according to the manufacturer’s recommendations.
Vujić T., Schvartz D., Iliuk A, & Sanchez J.C. (2021). Ubiquinone Metabolism and Transcription HIF-1 Targets Pathway Are Toxicity Signature Pathways Present in Extracellular Vesicles of Paraquat-Exposed Human Brain Microvascular Endothelial Cells. International Journal of Molecular Sciences, 22(10), 5065.
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5

Evaluating PQ Cytotoxicity in HBMEC

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HBMEC were seeded in a 96-well plate (10,000 cells per well) and treated for 24 h with PQ at different concentrations (0.1, 1, 10, 100, 1000 and 5000 µM). Cell proliferation was determined using the MTS assay (CellTiter 96 AQueous One Solution Cell Proliferation Assay, Promega), whereas cytotoxicity was assessed by measuring lactate dehydrogenase (LDH) released using a Pierce LDH cytotoxicity kit (Thermo Scientific). Both the MTS and LDH assays were performed according to the manufacturer’s recommendations.
Tatjana V., Domitille S, & Jean-Charles S. (2021). Paraquat-induced cholesterol biosynthesis proteins dysregulation in human brain microvascular endothelial cells. Scientific Reports, 11, 18137.
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6

Cytotoxicity Assay using LDH

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LDH release was measured using Pierce LDH cytotoxicity kit (Thermo Scientific, Pittsburgh, PA, USA) at 24 h after treatment. Absorbance was read at 450 μm using a Varioskan Flash Reader (Thermo Scientific, Waltham, MA, USA).
Liu X., Wen S., Yan F., Liu K., Liu L., Wang L., Zhao S, & Ji X. (2018). Salidroside provides neuroprotection by modulating microglial polarization after cerebral ischemia. Journal of Neuroinflammation, 15, 39.
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7

Crystal-Induced Cellular Cytotoxicity Assay

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BMMACs grown as described above were treated with indicated concentrations of crystals (MSU and Silica) and 10× lysis buffer (positive control). While cells were treated with the crystals for 5 hrs, 10× lysis buffer was given 45 minutes before sample collection. The supernatants were collected and LDH released was measured and calculated using Pierce LDH cytotoxicity kit from Thermo Scientific per manufacturer's instructions.
Hari A., Zhang Y., Tu Z., Detampel P., Stenner M., Ganguly A, & Shi Y. (2014). Activation of NLRP3 inflammasome by crystalline structures via cell surface contact. Scientific Reports, 4, 7281.
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8

Glomerular Endothelial Cell Assays

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Female C57BL/6 mice were purchased from The Taconic Laboratory. All experiments were performed in compliance with federal laws and institutional guidelines. The animal protocol was approved by the Augusta University Institutional Animal Care and Use Committee (no. A3307-01). 12-week-old mice (18–20 g) were used for all experiments. Established cloned glomerular endothelial cell clones were employed as described previously 28 (link). Urea nitrogen direct kit (Stanbio Laboratory, Boerne, TX), PKC-α inhibitor Ro-320432, (which displays 10-fold greator selectivity for PKC-α, 4-fold greater selectivity for PKC-β over other isoforms; EMD Millipore, Billerica, MA), PKC activator, Phorbol 12,13-dibutyrate (PDBu) (Sigma-Aldrich); EDC (Thermo Scientific, Rockford, IL), Mito-ID Membrane Potential Detection Kit (Enzo Life Sciences, Farmingdale NY), Pierce LDH Cytotoxicity Kit (Thermo Scientific), Nephrotoxicity PCR array (Qiagen, Maryland), MitoTracker Green FM (Thermo Scientific, Rockford, IL) wereused.
Kvirkvelia N., McMenamin M., Warren M., Jadeja R., Kodeboyina S.K., Sharma A., Zhi W., O’Conor P.M., Raju R., Lucas R, & Madaio M.P. (2018). Kidney targeted inhibition of protein kinase C-α ameliorates nephrotoxic nephritis with restoration of mitochondrial dysfunction. Kidney international, 94(2), 280-291.
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9

Astrocyte Proliferation and Cytotoxicity Assay

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Astrocytes were seeded in a 96-well plate (5000 cells per well) and treated for 24 h with TNF, IL-1β, or LPS. Cell proliferation was determined using the MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] assay (CellTiter 96® AQueous One Solution Cell Proliferation Assay, Promega), whereas cytotoxicity was assessed by measuring lactate dehydrogenase (LDH) release using a Pierce™ LDH cytotoxicity kit (Thermo Scientific). Both the MTS and LDH assays were performed according to the manufacturer’s recommendations.
Dozio V, & Sanchez J.C. (2018). Profiling the proteomic inflammatory state of human astrocytes using DIA mass spectrometry. Journal of Neuroinflammation, 15, 331.
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10

Morphine Cytotoxicity on HBMEC

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HBMEC were seeded in a 96-well plate (10,000 cells per well) and treated for 24 h with morphine at different concentrations (1, 10, 25, 50, 100, 200 and 400 µM). Cell proliferation was determined using the MTS assay (CellTiter 96®® AQueous One Solution Cell Proliferation Assay, Promega, Madison, WI, USA), whereas cytotoxicity was assessed by measuring lactate dehydrogenase (LDH) released using a Pierce™ LDH cytotoxicity kit (Thermo Scientific, Rockford, IL, USA). Both the MTS and LDH assays were performed according to the manufacturer’s recommendations.
Vujić T., Schvartz D., Furlani I.L., Meister I., González-Ruiz V., Rudaz S, & Sanchez J.C. (2022). Oxidative Stress and Extracellular Matrix Remodeling Are Signature Pathways of Extracellular Vesicles Released upon Morphine Exposure on Human Brain Microvascular Endothelial Cells. Cells, 11(23), 3926.
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