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Products:Kits/ Lysates/ Other >> Kits >> Cell Damage Kits
Overview
Product name
LDH-Cytotoxicity Assay Kit II (500 assays)
See all Lactate dehydrogenase products (3) ...
Assay type
Quantitative
Product overview
Cell death or cytotoxicity is classically evaluated by the quantification of plasma membrane damage. Lactate dehydragenase (LDH) is a stable enzyme, present in all cell types, and rapidly released into the cell culture medium upon damage of the plasma membrane. LDH, therefore, is the most widely used marker in cytotoxicity study. The LDH-Cytotoxicity Assay Kit II utilizes the advanced WST reagent for a fast and more sensitive detection of LDH released from damaged cells. The assay utilizing an enzymatic coupling reaction: LDH oxidizes lactate to generate NADH, which then reacts with WST to generate yellow color. The intensity of the generated color correlates directly with the cell number lysed. Since WST is brighter, less culture medium is required for the assay, and thus the background from serum and culture medium is significantly reduced. Using the assay, cells can be cultured in regular 10% serum containing medium, no reducing serum or special medium is required for the assay. In addition, since the WST is more stable, the reaction can be read multiple times, and can also be stopped at any time point during the reaction. LDH activity can be easily quantified by spectrophotometer or plate reader at OD450 nm. The kit provides all necessary reagents including LDH positive control. Assay takes less than 1 hour.
Tested applications
Functional Studiesmore details
Properties
Storage instructions
Store at -20°C, Stable for 6 months at -20°C
| Components | Identifier | 500 tests |
|---|---|---|
| Cell Lysis Solution | Clear | 1 x 6ml |
| LDH (0.1 µg/µl) | Red | 1 x 20µl |
| LDH Assay Buffer | NM | 1 x 50ml |
| Stop Solution | Clear | 1 x 6ml |
| WST Substrate Mix | Green | 1 vial |
Research areas
Kits/ Lysates/ Other >> Kits >> Cell Damage Kits >> Cell viability, plasma membrane damage
Cancer >> Cancer Metabolism >> Metabolic signaling pathway >> Metabolism of carbohydrates
Kits/ Lysates/ Other >> Kits >> Cell Metabolism Kits >> Sugar Assays
Kits/ Lysates/ Other >> Kits >> Cell Damage Kits
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Functional Studies - LDH-Cytotoxicity Assay Kit II (ab65393)
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Relevance
Lactate dehydrogenase (LDH) is an oxidoreductase which catalyses the interconversion of pyruvate and lactate with concomitant interconversion of NADH and NAD+. As it can also catalyze the oxidation of hydroxybutyrate, it is occasionally called Hydroxybutyrate Dehydrogenase (HBD). There are 5 different isoenzymes of LDH, LDH1 to LDH5, each composed of 4 subunits which may be of 2 different types - M and H subunits. These subunits are encoded by two different genes: The M subunit is encoded by gene LDHA whilst the H subunit is encoded by LDHB. Usually LDH2 is the predominant form in the serum. An LDH1 level higher than the LDH2 level suggests myocardial infarction (damage to heart tissues releases heart LDH, which is rich in LDH1, into the bloodstream).
Cellular localization
Cytoplasmic
Alternative names
- L lactate dehydrogenase A chain
- L lactate dehydrogenase B chain
- Lactate dehydrogenase A
see all
Applications
Show applications key
Our Abpromise guarantee covers the use of ab65393 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
FuncS: Use at an assay dependent dilution.
LDH-Cytotoxicity Assay Kit II (500 assays) images:
Functional Studies - LDH-Cytotoxicity Assay Kit II (ab65393)
Comparison of WST-1 and INT based assays. 3T3 cells were cultured in a 96-well plate in 100 µl of culture medium. LDH assays were performed using 10 µl of culture medium using WST-1 (Brown bar) and INT (Green bar) methods. The WST-1 based assay is more stable and sensitive than the INT based method.
Protocols
References for LDH-Cytotoxicity Assay Kit II (500 assays) (ab65393)
This product has been referenced in:
- Petrasek Jet al. Type I interferons protect from Toll-like receptor 9-associated liver injury and regulate IL-1 receptor antagonist in mice. Gastroenterology 140:697-708.e4 (2011). Mouse.Read more (PubMed: 20727895) »
- Wang Ket al. Export of microRNAs and microRNA-protective protein by mammalian cells. Nucleic Acids Res 38:7248-59 (2010). Human.Read more (PubMed: 20615901) »
See all 2 publications for this product
Publishing research using ab65393? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"