SH-SY5Y cells were transfected with human ClpP siRNA or control siRNA for 3 days. After then, the cells were cultured in FBS-free DMEM/F12 (1:1) medium for 24 h. Neurons derived from iPS cells carrying αSyn A53T and its corrected isogenic control were cultured in neuronal differentiation medium (GDNF, ascorbic acid, TGFβ, cAMP) without BDNF for 24 h. Cell death was determined by measuring LDH release into the culture medium, using LDH-Cytotoxicity Assay Kit II (Roche, USA, 04744926001) by following the manufacturer’s instruction.
Ldh cytotoxicity assay kit 2
Manufactured by Roche
Sourced in United States
The LDH-Cytotoxicity Assay Kit II is a colorimetric assay used to measure lactate dehydrogenase (LDH) activity released from damaged cells. It provides a quantitative assessment of cytotoxicity or cell death in cell culture systems.
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Lab products found in correlation
6 protocols using ldh cytotoxicity assay kit 2
1
SH-SY5Y and iPSC Neuron Cell Death Assay
2
Investigating HMGB1 Release in Cell Lines
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HdhQ7 and Q111 mouse striatal cells were treated with the HV-3 peptide or the control peptide TAT (3 μM, each) in an FBS-free DMEM medium or in DMEM containing 10% serum for 24 h. Medium from the cultured cells was harvested. Proteins from the medium were purified using Amicon Ultra 0.5 ml centrifugal filters (Millipore). HMGB1 release into the medium was then analysed by Western blotting with anti-HMGB1 antibody. In parallel, cell death was determined by measuring LDH release into the culture medium, using LDH-Cytotoxicity Assay Kit II (Roche, USA) by following the manufacturer's instruction.
3
Assessing Cell Death in Neurodegeneration
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SH-SY5Y cells were transfected with human ClpP siRNA or control siRNA for 3 days. After then, the cells were cultured in FBS-free DMEM/F12 (1:1) medium for 24 h. Neurons derived from iPS cells carrying αSyn A53T and its corrected isogenic control were cultured in neuronal differentiation medium (GDNF, ascorbic acid, TGFβ, cAMP) without BDNF for 24 h. Cell death was determined by measuring LDH release into the culture medium, using LDH-Cytotoxicity Assay Kit II (Roche, USA, 04744926001) by following the manufacturer’s instruction.
4
Cell Membrane Integrity Assessment
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The cell membrane integrity was measured using a LDH Cytotoxicity Assay Kit II (Roche, Grenzach, Germany) according to the manufacturer’s instructions. Briefly, 10 µL of cell culture supernatant was added to 100 µL of LDH reaction mix, and incubated for 30 min at room temperature in the dark. The absorbance was measured at 450 nm (reference wavelength: 650 nm) using a photometer (Tecan infinite M200 pro). The cell membrane integrity was normalized to the number of vital adherent cells per mm2 (n = 36; three independent experiments with three different wells per experiment, measured in four technical replicates).
5
Cytotoxicity Measurement by LDH Assay
6
Striatal Cell Death Assay
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Mouse striatal HdhQ7/HdhQ111 cells were treated with 3 µM CHIR99021 for 2 days and then serum-starved for 16 h. Cell death was determined by measuring LDH release into the medium using an LDH-cytotoxicity assay kit II (Roche) following the manufacturer’s instructions. Relative LDH release was calculated as: LDHmedium/(LDHmedium + LDHcell) and then normalized to HdhQ7 cells data.
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