Product nameLDH Assay Kit (Cytotoxicity)
See all Lactate Dehydrogenase kits
Sample typeAdherent cells, Suspension cells
Assay typeEnzyme activity (quantitative)
Assay time1h 00m
LDH Assay Kit (Cytotoxicity) ab65393 (previously called LDH-Cytotoxicity Assay Kit II) uses WST reagent for a fast and more sensitive detection of LDH released from damaged cells.
The LDH assay protocol uses an enzymatic coupling reaction: LDH oxidizes lactate to generate NADH, which then reacts with WST to generate a yellow color. The intensity of the generated color correlates directly with the cell number lysed.
As WST is brighter than other cell viability reagents, less culture medium is required for the assay, and thus the background from serum and culture medium is significantly reduced. Cells can be cultured in regular 10% serum containing medium; no reducing serum or special medium is required for the assay.
In addition, since WST is very stable, the reaction can be read multiple times and can be stopped at any time point during the reaction.
LDH activity can be easily quantified by spectrophotometer or plate reader at OD450nm.
LDH assay protocol summary:
- transfer culture medium into new plate
- add LDH reaction mix and incubate for 30 min at room temp
- analyze with microplate reader
If you would like to use a fluorometric reading, please refer to LDH-Cytotoxicity Assay Kit (Fluorometric) (ab197004).
This kit is more sensitive than our colorimetric LDH Cytoxicity Assay Kit ab65391.
A cell death assay / cytotoxicity assay classically assesses the level of plasma membrane damage to a cell population. Lactate dehydrogenase (LDH) is a stable enzyme, present in all cell types, which is rapidly released into the cell culture medium upon damage of the plasma membrane. LDH is the most widely used marker used to run a cytotoxicity assay.
Storage instructionsStore at -20°C. Please refer to protocols.
Components Identifier 500 tests Cell Lysis Solution Clear 1 x 5ml LDH (Positive control) Red 1 vial LDH Assay Buffer NM 1 x 50ml Stop Solution Blue 1 x 5ml WST Substrate Mix Amber 1 vial
- Pathways and Processes
- Metabolic signaling pathways
- Energy transfer pathways
- Energy Metabolism
PathwayFermentation; pyruvate fermentation to lactate; (S)-lactate from pyruvate: step 1/1.
Involvement in diseaseDefects in LDHA are the cause of glycogen storage disease type 11 (GSD11) [MIM:612933]. A metabolic disorder that results in exertional myoglobinuria, pain, cramps and easy fatigue.
Sequence similaritiesBelongs to the LDH/MDH superfamily. LDH family.
- Information by UniProt
- Cell proliferation-inducing gene 19 protein
- L lactate dehydrogenase B chain
Bafilomycin A1 (BafA1) toxicity was assessed in Human hepatic (Huh7.5) cells after 24 hours at different concentrations (12.5nm, 25nm, 50nm and 100nm) were administered to cells using LDH cytotoxicity assay kit (ab65393). Cytotoxicity was measured by subtracting LDH content in remaining viable cells from total LDH in untreated controls.
Jurkat T cells were cultured in 96-well plate in 100 μl of culture medium. LDH Assay was performed using 10μl of culture medium using the WST probe. Low control (white bar); High control (black bar).
Comparison of WST-1 and INT based LDH assays. 3T3 cells were cultured in a 96-well plate in 100 µl of culture medium. The LDH assay was performed using 10 µl of culture medium using WST-1 (Brown bar) and INT (Green bar) methods. The WST-1 based LDH assay is more stable and sensitive than the INT based method.
This product has been referenced in:
- Jackson TC et al. BrainPhys® increases neurofilament levels in CNS cultures, and facilitates investigation of axonal damage after a mechanical stretch-injury in vitro. Exp Neurol 300:232-246 (2018). Functional Studies . Read more (PubMed: 29199132) »
- Smith JA et al. RNA Nanotherapeutics for the Amelioration of Astroglial Reactivity. Mol Ther Nucleic Acids 10:103-121 (2018). Functional Studies . Read more (PubMed: 29499926) »