Key features and details
- Assay type: Enzyme activity (quantitative)
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 1 hr
- Sample type: Cell culture media
Product nameLDH Assay Kit (Cytotoxicity)
See all Lactate Dehydrogenase kits
Sample typeCell culture media
Assay typeEnzyme activity (quantitative)
Assay time1h 00m
LDH Assay Kit (Cytotoxicity) ab65393 uses WST for the fast and sensitive detection of LDH released from damaged cells.
The LDH assay, also known as LDH release assay, is a cell death / cytotoxicity assay used to assess the level of plasma membrane damage in 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.
The LDH assay protocol is based on an enzymatic coupling reaction: LDH released from the cell 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 number of lyzed cells.
Only 10µl of 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 10µl culture medium into new plate
- add LDH reaction mix and incubate for 30 min at room temp
- analyze with microplate reader
This kit was previously called LDH-Cytotoxicity Assay Kit II.
Alternative LDH assays
If you would like to use a fluorometric assay, please refer to LDH-Cytotoxicity Assay Kit (Fluorometric) (ab197004).
This kit is more sensitive than the colorimetric LDH Cytoxicity Assay Kit ab65391.
To measure LDH activity in sample types such as serum, plasma, and cell lysates, we recommend LDH assay kit ab102526.
Related products and guides to cytotoxicity / cell viability / proliferation assays
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Storage instructionsStore at -20°C. Please refer to protocols.
Components Identifier 500 tests Cell Lysis Solution Clear/NM 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.
ab65393 has been referenced in 59 publications.
- Song N et al. Heat Shock Protein 70 Protects the Heart from Ischemia/Reperfusion Injury through Inhibition of p38 MAPK Signaling. Oxid Med Cell Longev 2020:3908641 (2020). PubMed: 32308802
- Wei KC et al. Assessment of Prolonged Dengue Virus Infection in Dermal Fibroblasts and Hair-Follicle Dermal Papilla Cells. Viruses 12:N/A (2020). PubMed: 32121148
- Ahmed F et al. The effects of acute BPA exposure on skeletal muscle mitochondrial function and glucose metabolism. Mol Cell Endocrinol 499:110580 (2020). PubMed: 31536778
- Lim D et al. Targeted Delivery of the Mitochondrial Target Domain of Noxa to Tumor Tissue via Synthetic Secretion System in E. coli. Front Bioeng Biotechnol 8:840 (2020). PubMed: 32766235
- Carey RM et al. Polarization of protease-activated receptor 2 (PAR-2) signaling is altered during airway epithelial remodeling and deciliation. J Biol Chem 295:6721-6740 (2020). PubMed: 32241907