Description
Key facts
Sample types:Suspension cells, Adherent cells,
Assay type:Enzyme activity,
Assay time:1h,
Assay Platform:Microplate reader
Product details:
For a more sensitive LDH Assay Kit to use as a cytotoxicity assay, we recommend
ab65393
LDH Assay Kit (Cytotoxicity) ab65391 provides a fast and simple method for quantitating cytotoxicity based on the measurement of activity of lactate dehydrogenase (LDH) released from damaged cells.
The assay takes ~0.5-1 hr. LDH activity can be determined by a coupled enzymatic reaction: LDH oxidizes lactate to pyruvate which then reacts with tetrazolium salt INT to form formazan. The increase in the amount of formazan produced in culture supernatant directly correlates to the increase in the number of lysed cells. The formazan dye is water-soluble and can be detected by spectrophotometer at 500 nm.
This kit was previously called LDH-Cytotoxicity Assay Kit I.
Other Notes
Cell death or cytotoxicity is classically evaluated by the quantification of plasma membrane damage. Unlike many other cytoplasmic enzymes which exist in many cells either in low amount (e.g., alkaline and acid phosphatase) or unstable, LDH is a stable cytoplasmic enzyme present in all cells and rapidly released into the cell culture supernatant upon damage of the plasma membrane.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Lactate dehydrogenase (LDH) is an enzyme that catalyzes the interconversion of pyruvate and lactate along with the conversion of NADH to NAD+. LDH is known by other names such as lactic acid dehydrogenase and LDH-5. The enzyme has a molecular weight of approximately 36 kDa. LDH exists in almost all tissues having multiple isoforms that are expressed differently depending on the tissue type. It shows high expression in muscle tissue liver and heart indicating its extensive role in energy metabolism.
Biological function summary
Lactate dehydrogenase plays a critical role in anaerobic glycolysis. The enzyme helps in regenerating NAD+ from NADH allowing glycolysis to continue in the absence of oxygen. LDH is not a part of any larger protein complex working independently to fulfill its function in the glycolytic pathway. It serves in rapid energy production especially under hypoxic or exertional conditions where oxygen supply is limited.
Pathways
LDH is significantly involved in the glycolysis and gluconeogenesis pathways. Within glycolysis LDH helps facilitate the conversion of pyruvate to lactate during anaerobic conditions a step important for ATP production when oxygen is scarce. The enzyme is tied closely to phosphofructokinase-1 (PFK-1) in glycolysis given that both enzymes are central to maintaining the glycolytic flow. In gluconeogenesis though functionally reversed from its role in glycolysis LDH helps to manage lactate removal an important step for glucose synthesis from non-carbohydrate sources.
Lactate dehydrogenase levels often act as a biomarker for tissue damage or certain cancers as its release into the bloodstream signals cellular injury or death. Elevated LDH levels are associated with conditions like myocardial infarction and certain forms of anemia. In cancer such as lymphoma or leukemia LDH correlates with the progression of the disease and acts as a prognostic marker. LDH's connection to these conditions often leads to insights into disease severity and progression due to its association with proteins like p53 and HIF-1 which play roles in cellular metabolism and hypoxia response.
