Cytotoxicity Detection Kit^PLUS (LDH)

Nonradioactive colorimetric assay suitable for high-throughput quantification of cell death and cell lysis, based on the measurement of lactate dehydrogenase (LDH) activity released from the cytosol of damaged cells into the supernatant. The Cytotoxicity Detection Kit^PLUS (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.

• The Cytotoxicity Detection Kit^PLUS (LDH) is a fast, sensitive, and simple method to quantify 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.

The assay measures LDH activity released from damaged cells. It is performed in 96-well plates.

• Flexible, since the assay can determine the degree of plasma membrane damage in many different in vitro cell systems.
• Safe, because the assay does not use radioactive isotopes.
• Accurate, since assay results strongly correlate to the number of lysed cells.
• Sensitive, since the assay can detect even low cell numbers.
• Fast, since assay results can be determined on a multi-well ELISA reader, which allows simultaneous processing of many samples.
• Convenient, since the assay does not require a prelabeling step.

1 kit containing 4 components

Working solution: Catalyst:

Dissolve the lyophilizate in 1 ml double dist. water for 10 min, then mix thoroughly.

Reaction mixture:

For 100 tests: Shortly before use, mix 250 μl of reconstituted catalyst with 11.25 ml dye solution
For 400 tests: Shortly before use, add the total volume of reconstituted catalyst (1 ml) to the total volume of dye solution (45 ml) and mix well.
Note: In case you observe crystals in “Dye solution”, shake the bottle at least 1 hour at 37 °C. Remaining precipitates will NOT influence the performance.
Note: The precipitates were formed during the freezing of the product. Repeated freezing and thawing should be avoided.
Note: Prepare immediately before use. Do not store the Reaction mixture.
Storage conditions (working solution): The lyophilizate is stable at 2 to 8 °C.
The reconstituted catalyst solution, the dye solution the lysis solution and the stop solution are stable for

• Four weeks when stored at 2 to 8 °C
• Two days at 15 to 25 °C
• For long-term storage (up to 3 months), store bottles at -15 to -25 °C.
• Freezing and thawing the solutions up to three times will not significantly reduce their performance. When crystals are seen in Dye solution, shake the bottle at least one hour at 37 °C. Remaining precipitates will not influence performance. As the precipitates can be formed during freezing of the product, repeated freezing and thawing should be avoided.

For life science research only. Not for use in diagnostic procedures.