Cytotoxicity Chromogenic Assay Kit from Takara Bio USA

Description

Takara’s LDH Cytotoxicity Detection Kit allows simple measurement of LDH activity released from the of damaged cells into the supernatant. This kit is designed to allow a precise, fast and simple colorimetric assay method to quantitate cytotoxicity/cytolysis based on the measurement of LDH activity released from damaged cells into the supernatant. This is a non-radioactive alternative assay method to the [3H]-thymidine release assay and the [51Cr]-release assay. The culture supernatant is collected cell-free and incubated with the reaction mixture from the kit, and the LDH activity is determined in an enzymatic test as below: In the first step NAD+ is reduced to NADH/H+ by the LDH-catalyzed conversion of lactate to pyruvate. In the second step, the catalyst (diaphorase) transfers H/H+ from NADH/H+ to the tetrazolium salt INT which is reduced to formazan.An increase in the number of dead or plasma membrane-damaged cells leads to an increase of the LDH enzyme activity in the culture supernatant. This increase of the enzyme activity in the supernatant directly correlates to the amount of formazan formed in a certain time period. That is, the amount of color formed in the assay is proportional to the number of damaged cells. The formazan dye formed is water soluble and shows the maximum absorption at about 500 nm, whereas the tetrazolium salt INT shows no significant absorption at these wave lengths

Applications

• Detection and quantification of cell mediated cytotoxicity induced by cytotoxic T-lymphocytes (CTL), natural killer (NK) cells, lymphokine activated killer (LAK) cells or monocytes
• Determination of mediator which induces cytolysis
• Measurement of antibody-dependent cellular cytotoxicity (ADCC) and complement-mediated cytolysis
• Determination of the cytotoxic potential of compounds in environmental and medical research and in the food, cosmetic and pharmaceutical manufacturing
• Determination of cell death in bioreactors

Features

• Safe: No radioactive isotopes are used.
• Accurate: Assay results obtained with this kit strongly correlate to the number of damaged cell.
• High sensitivity: Low cell numbers, such as 0.2 - 2 x 10² cells/well, can be available for detection.
• Fast: A large number of samples can be processed simultaneously by using a multiwell-ELISA reader. It takes only 0.5 - 1 hour for measurement.
• Simple procedure: - No need for prelabeling and washing steps. - As this kit does not employ radioactive isotopes, no disposal and radiation safety operations or paperwork are required.
• Guaranteed performance: Performance is tested with every lot.