LDH Assay Kit ab65393 is a no-wash assay with one incubation step of 30 min at room temp. LDH, in cell culture supernatant samples, oxidizes lactate to generate NADH. NADH reacts with WST, with a colorimetric readout (450 nm). The LDH signal is proportional to the amount of cytotoxicity.
Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
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Publications
- Molecular psychiatry 29:369-3862023Cerebral organoids with chromosome 21 trisomy secrete Alzheimer's disease-related soluble aggregates detectable by single-molecule-fluorescence and super-resolution microscopy.Applications:Unspecified applicationReactive species:Unspecified reactive speciesEmre Fertan,Dorothea Böken,Aoife Murray,John S H Danial,Jeff Y L Lam,Yunzhao Wu,Pollyanna A Goh,Ivan Alić,Matthew R Cheetham,Evgeniia Lobanova,Yu P Zhang,Dean Nižetić,David KlenermanPubMed 38102482
- mBio 14:e01979232023Enteropathogenic infection co-elicits lysosomal exocytosis and lytic host cell death.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRaisa Shtuhin-Rahav,Aaron Olender,Efrat Zlotkin-Rivkin,Etan Amse Bouman,Tsafi Danieli,Yael Nir-Keren,Aryeh M Weiss,Ipsita Nandi,Benjamin AroetiPubMed 38038448
Key facts
- Detection method
Colorimetric
- Sample types
Cell culture media
- Assay type
Enzyme activity (quantitative)
- Assay time
1h
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Alternative names
L-lactate dehydrogenase A chain, LDH-A, Cell proliferation-inducing gene 19 protein, LDH muscle subunit, Renal carcinoma antigen NY-REN-59, LDH-M, PIG19, LDHA
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LDH Assay Kit ab65393 is a no-wash assay with one incubation step of 30 min at room temp. LDH, in cell culture supernatant samples, oxidizes lactate to generate NADH. NADH reacts with WST, with a colorimetric readout (450 nm). The LDH signal is proportional to the amount of cytotoxicity.
Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
Alternative names
L-lactate dehydrogenase A chain, LDH-A, Cell proliferation-inducing gene 19 protein, LDH muscle subunit, Renal carcinoma antigen NY-REN-59, LDH-M, PIG19, LDHA
Key facts
- Detection method
Colorimetric
- Sample types
Cell culture media
- Assay type
Enzyme activity (quantitative)
- Assay time
1h
- Assay Platform
Microplate reader
Storage
- Shipped at conditions
Blue Ice
- Appropriate short-term storage conditions
-20°C
- Appropriate long-term storage conditions
-20°C
- Storage information
-20°C
Notes
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 . This kit is more sensitive than the .To measure LDH activity in sample types such as serum, plasma, and cell lysates, we recommend .**Related products and guides to cytotoxicity / cell viability / proliferation assays**Review our to learn about our other kits to perform a , or .
Supplementary info
- Activity summary
Lactate dehydrogenase (LDH) also known as LD or LDH1 is an enzyme that plays an important role in the conversion between lactate and pyruvate. It catalyzes the oxidation of L-lactate to pyruvate while simultaneously reducing NAD+ to NADH. LDH is approximately 140 kDa in size and consists of tetrameric structures formed by different combinations of its subunits. Expression of LDH occurs in many tissues with significant presence in muscles liver kidneys and the heart.
- Biological function summary
The enzyme works efficiently to help cells convert glucose into energy under conditions lacking oxygen such as intense exercise or hypoxia. LDH does not act alone; it functions as part of a larger metabolic network where it works with other enzymes and proteins to maintain cellular energy balance. The enzyme helps in maintaining the redox balance within the cell by regulating levels of NAD+ and NADH. Due to its importance in metabolism LDH is often measured in laboratories using lactate dehydrogenase assays including LDH release assays and LDH activity assays.
- Pathways
LDH is an integral part of the glycolysis and gluconeogenesis pathways. It facilitates the conversion of pyruvate to lactate especially during anaerobic glycolysis enabling the continual production of ATP in oxygen-poor environments. LDH is associated with other metabolic proteins like pyruvate kinase and hexokinase each playing their own roles in energy production processes. Understanding these interactions is important in research that involves energy metabolism.
- Associated diseases and disorders
LDH levels can indicate tissue damage or disease progression. Elevated LDH levels are often associated with conditions such as myocardial infarction and various types of cancers. For example in cancer LDH levels increase due to high anaerobic glycolysis rates commonly observed in tumor cells. LDH also relates to diagnostic markers in myocardial infarctions where it increases in response to cardiac tissue damage. LDH assays are frequently used as part of diagnostic protocols to assess the severity or presence of such conditions.
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3 product images
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/Functional Studies - LDH Assay Kit (Cytotoxicity) (ab65393)
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.
Functional Studies - LDH Assay Kit (Cytotoxicity) (ab65393)
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).
Functional Studies - LDH Assay Kit (Cytotoxicity) (ab65393)
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.
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