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Ldh cytotoxicity detection assay

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The LDH (lactate dehydrogenase) cytotoxicity detection assay is a colorimetric method used to quantify cell death or lysis. The assay measures the release of LDH, a stable cytoplasmic enzyme, from damaged cells. The amount of LDH released is proportional to the number of lysed or damaged cells, which can be used as an indicator of cytotoxicity.

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Lab products found in correlation

Trypan blue, by Merck Group (1 mentions) Dispase, by Thermo Fisher Scientific (1 mentions) Tesr e8 medium, by STEMCELL (1 mentions) Matrigel, by STEMCELL (1 mentions) Gentle cell dissociation reagent (gcdr), by STEMCELL (1 mentions) Matrigel, by BD (1 mentions) Tc20 counter, by Bio-Rad (1 mentions)

Close spelling variants of this product

LDH cytotoxicity assay Cytotoxicity detection assay LDH assay

3 protocols using ldh cytotoxicity detection assay

1

LDH Cytotoxicity Assay for Lung Tissue

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Tissue samples were treated with 10 uM BI2536 compound, DMSO or 0.2% triton for 48 h. Standard lactate dehydrogenase (LDH) cytotoxicity detection assay (Roche) was performed with supernatants (100 μl) according to manufacturer’s recommendations. For data analysis, LDH activity at 0.2% triton was used as positive control and set as 1. Toxicity was measured as LDH activity relative to positive control.
All experiments using human lung tissue samples were carried out in accordance with relevant guidelines and regulations according to the Cooperative Human Tissue Network (www.chtn.org). The experimental protocols were approved by the Sanford Burnham Prebys Medical Discovery Institute Institutional Review Board (IRB). Informed consent was obtained from all subjects.
Pohl M.O., von Recum-Knepper J., Rodriguez-Frandsen A., Lanz C., Yángüez E., Soonthornvacharin S., Wolff T., Chanda S.K, & Stertz S. (2017). Identification of Polo-like kinases as potential novel drug targets for influenza A virus. Scientific Reports, 7, 8629.
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2

Titanium Disc Cell Cytotoxicity Assay

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Cells were sowed over titanium discs. They were incubated for 48 h and then, stimulated as previously described. After 24 h at 37 °C in a 5% CO2 incubator, the lactate dehydrogenase (LDH) cytotoxicity detection assay (Roche, Basel, Switzerland) was carried out according to the manufacturer’s instructions.
González-Sánchez Z., Areal-Quecuty V., Jimenez-Guerra A., Cabanillas-Balsera D., Gil F.J., Velasco-Ortega E, & Pozo D. (2022). Titanium Surface Characteristics Induce the Specific Reprogramming of Toll-like Receptor Signaling in Macrophages. International Journal of Molecular Sciences, 23(8), 4285.
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3

Culturing Human Embryonic Stem Cells

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Human ESCs (H9 (WA09); passages 30–50) were obtained from the WiCell Research Institute (Madison, WI). Cells cultured in dishes coated with Matrigel (BD Biosciences, San Jose, CA) or recombinant human vitronectin, truncated (VN; cat. no. A14700, Life Technologies, Grand Island, NY) and in TeSR-E8 medium (E8; Stem Cell Technologies, Vancouver, BC) were maintained in 5% CO2/95% air at 37 °C. Medium was replaced every day, and the cells were passaged every 5–6 days by enzymatic dissociation with dispase (Life Technologies, Grand Island, NY) for cells cultured on Matrigel or with Gentle Cell Dissociation reagent (Stem Cell Technologies).
Viable cells were counted in a hemocytometer or using a TC-20 counter (Bio-Rad, Hercules, CA) after Trypan Blue staining (Sigma-Aldrich, St. Louis, MO). Alternatively, cells were stained with 20 μg/ml fluorescein diacetate (FDA-live cells; Sigma-Aldrich) in PBS for 5 min and after being washed twice with PBS, they were analyzed by fluorescence microscopy or flow cytometry.
Lactate dehydrogenase (LDH) activity was determined in culture samples with a LDH cytotoxicity detection assay (Roche, Indianapolis, IN) according to the manufacturer’s instructions2 (link), 13 (link).
Fan Y., Zhang F, & Tzanakakis E.S. (2016). Engineering Xeno-Free Microcarriers with Recombinant Vitronectin, Albumin and UV Irradiation for Human Pluripotent Stem Cell Bioprocessing. ACS biomaterials science & engineering, 3(8), 1510-1518.
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