LDH Assay Kit (Cytotoxicity) (ab65393)
Key features and details
- Assay type: Enzyme activity (quantitative)
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 1 hr
- Sample type: Cell culture media
Overview
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Product name
LDH Assay Kit (Cytotoxicity)
See all Lactate Dehydrogenase kits -
Detection method
Colorimetric -
Sample type
Cell culture media -
Assay type
Enzyme activity (quantitative) -
Assay time
1h 00m -
Product overview
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 -
Notes
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
Review our cell health assay guide to learn about our other kits to perform a cell viability assay, cytotoxicity assay or cell proliferation assay.
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. -
Platform
Microplate reader
Properties
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Storage instructions
Store 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 -
Research areas
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Pathway
Fermentation; pyruvate fermentation to lactate; (S)-lactate from pyruvate: step 1/1. -
Involvement in disease
Defects 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 similarities
Belongs to the LDH/MDH superfamily. LDH family. -
Post-translational
modificationsISGylated. -
Cellular localization
Cytoplasm. - Information by UniProt
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Alternative names
- Cell proliferation-inducing gene 19 protein
- GSD11
- L lactate dehydrogenase B chain
see all
Images
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LDH Cytotoxicity Assay using ab65393 Image from Bukong TN et al., PLoS Pathog 10(10), Fig S6. Doi: 10.1371/journal.ppat.1004424. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
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.
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LDH Cytotoxicity Assay useing ab65393Jurkat 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).
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LDH Cytotoxicity Assay using ab65393Comparison 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.
