LDH-Cytotoxicity Assay Kit provides a fast and simple method for quantitating cytotoxicity based on the measurement of activity of lactate dehydrogenase (LDH) released from damaged cells. The LDH-cytotoxicity assay is sensitive, convenient, and precise, and is applicable to a variety of cytotoxicity studies. 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.
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 LDH-Cytotoxicity Assay Kit (Fluorometric).
Store kit at -20°CCATALYST SOLUTION: Reconstitute the Catalyst in 1 ml ddH2O for 10 min and mix thoroughly. The Catalyst Solution is Stable for several weeks at +4°C.DYE SOLUTION: After thawing, the Dye Solution is Stable for several weeks at +4°C. Avoid freeze/thaw cycles.REACTION MIXTURE: For 100 assays, mix 250 μL of Catalyst Solution with 11.25 ml of Dye Solution. The mixture solution should be prepared immediately before use.
Components 400 testsCatalyst (Lyophilized) 1 vialDye Solution 1 x 45ml
Compatible Sample Types:
Adherent cells, Suspension cells