LDH Cytotoxicity BioAssay(TM) Kit II (Colorimetric)

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. Sample Type: Cell culture (adherent and suspension) Intended Use: This assay provides a convenient and Wst-based non-radioactive assay for measurement of activity of lactate dehydrogenase (LDH) which is a stable enzyme normally found in the cytosol of all cells but rapidly releases into the supernatant upon damage of plasma membrane.? Test Principle: LDH Cytotoxicity BioAssay(TM) 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 amount of 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. Kit Components: WST Substrate Mix, 1x vial LDH Assay Buffer, 1x50ml Cell Lysis Solution, 1x6ml Stop Solution, 1x6ml LDH, 1x20ul Storage and Stability: Store powder at 4 degrees C liquid at -20 degrees C. Store other components at 4 degrees C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.