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LDH Assay Kit ab65391 provides a fast and simple method cytotoxicity assay based on measurement of activity of LDH released from damaged cells.For a more sensitive LDH Assay Kit to use as a cytotoxicity assay, we recommend ab65393.

Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.

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Images

Functional Studies - LDH Assay Kit (Cytotoxicity) (AB65391), expandable thumbnail
  • Functional Studies - LDH Assay Kit (Cytotoxicity) (AB65391), expandable thumbnail

Publications

Key facts

Sample types

Suspension cells, Adherent cells

Assay type

Enzyme activity

Assay time

1h

What's included?

8000 Test
Components
Dye Solution I
20 x 45 mL
LDH Catalyst Reagent
20 x 1 Vial

Recommended products

LDH Assay Kit ab65391 provides a fast and simple method cytotoxicity assay based on measurement of activity of LDH released from damaged cells.For a more sensitive LDH Assay Kit to use as a cytotoxicity assay, we recommend ab65393.

Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.

Key facts

Sample types

Suspension cells, Adherent cells

Assay type

Enzyme activity

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

For a more sensitive LDH Assay Kit to use as a cytotoxicity assay, we recommend LDH Assay Kit (Cytotoxicity) 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.

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. If you would like to use a fluorometric reading, please refer to .

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

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.

Associated diseases and disorders

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.

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2 product images

  • Functional Studies - LDH Assay Kit (Cytotoxicity) (ab65391), expandable thumbnail

    Functional Studies - LDH Assay Kit (Cytotoxicity) (ab65391)

    LDH release measured at various cell amounts

  • Functional Studies - LDH Assay Kit (Cytotoxicity) (ab65391), expandable thumbnail

    Functional Studies - LDH Assay Kit (Cytotoxicity) (ab65391)

    Jurkat cells were cultured in a 96-well plate in 100 µl of culture medium. LDH assay was performed using 10 µl of culture medium according to the kit instructions. Light bar: Low control; Dark bar: High control.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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