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Cytotoxicity Detection KitPLUS (LDH)
Application
The Cytotoxicity Detection KitPLUS (LDH) is a fast, sensitive, and simple method to quantitate cytotoxicity/cytolysis based on the measurement of LDH activity released from damaged cells using the 96-well or 384-well plate format. The kit can be used in many different in vitro cell systems when damage to the plasma membrane occurs. For example:
Detection and quantification of cell-mediated cytotoxicity.
Determination of mediator-induced cytolysis.
Determination of the cytotoxic potential of compounds in environmental and medical research, and in the food, cosmetic, and pharmaceutical industries.
Determination of cell death in bioreactors.
Benefits
Suitable for high throughput: Fewer handling steps. No transfer, centrifugation, or prelabeling steps are required.
Flexible: Defined assay conditions when using a stopped color reaction.
Safe: No radioactive isotopes are used.
Accurate: Results obtained strongly correlate to lysed cell number.
Sensitive: Low cell numbers are detected.
Fast: The use of a multiwell ELISA reader permits a larger number of samples to be processed.
Product Description
The Cytotoxicity Detection KitPLUS (LDH) can be performed in a homogeneous format and requires no transfer and centrifugation steps to separate the supernatant from the cells. The color reaction can be stopped for defined assay conditions.
Sample material: Cell cultures grown in 96- or 384-well plates can be measured directly. Aliquots from cultures grown in other formats can be transferred into 96- or 384-well plates for measurement without removing the cells.
Assay time: 10 to 30 minutes for incubation.
Sensitivity: Less than 100 lysed cells can be detected in a 96-well plate.
Background Information
Cell death is classically evaluated by the quantification of plasma-membrane damage. Widely used methods are based on the differential uptake or exclusion of dyes, such as trypan blue and propidium iodide, followed by counting using a microscope. The major disadvantage is that these methods do not permit processing of large sample numbers. Another group of assays is based on the release of radioactive isotopes such as [51Cr] and [3H]-thymidine or fluorescence dyes from prelabeled target cells. The disadvantages of these assays are (i) the use of radioactive isotopes in most of them, (ii) the necessity for prelabeling of the target cells, and (iii) the high spontaneous release of most labels from the prelabeled target cells.
A third type of assay is based on the measurement of cytoplasmic enzyme activity released by damaged cells. Enzyme-release assays have been described (e.g., for alkaline and acid phosphatase); however, their use is hampered by the low amount of relevant enzymes that is present in many cells, and by the elaborate kinetic assays required to quantitate most enzyme activities. In contrast, lactate dehydrogenase (LDH) is a stable cytoplasmic enzyme that is present in all cells. It is rapidly released into the cell-culture supernatant upon damage of the plasma membrane.
Contents
Catalyst (Diaphorase/NAD+ mixture)
Dye Solution (INT and sodium lactate)
Lysis Solution
Stop Solution
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