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LDH-Cytotoxicity Assay Kit (Fluorometric) (ab197004) provides a sensitive, quick, and easy way to detect LDH (lactate dehydrogenase) released from damaged cells. In this assay, LDH converts lactate to pyruvate and NADH, which reduces a proprietary probe to an intensely fluorescent product (Ex/Em = 535/587 nm). The amount of fluorescence is directly proportional to the number of damaged cells. The assay is adaptable to high-throughput format and can be completed in less than 20 min. Sensitivity: ~ 100 cells.
The protocol of this product has been optimized for use on 96-well tissue culture plate, but it can be easily modified to be used with 6-, 12- or 24-well tissue culture plates.
Cell death or cytotoxicity is classically evaluated by the quantification of plasma membrane damage. Lactate dehydragenase (LDH, EC 18.104.22.168) is a stable enzyme, present in all cell types, which is rapidly released into the cell culture medium upon damage of the plasma membrane. LDH, therefore, is the most widely used marker in cytotoxicity study.
|Cell Lysis Solution||1 x 5ml|
|LDH Assay Buffer||1 x 50ml|
|LDH Substrate Mix||Amber||1 vial|
|PicoProbe||Blue||1 x 2ml|
|Positive Control||1 x 20µl|
Untreated Jurkat cells (low control) or treated with Cell Lysis Solution for 30 min (high control).
Relative fluorescence units of untreated Jurkat cells (low control) or treated with Cell Lysis Solution for 30 min (high control).
Overnight treatment of HeLa cells with 100μM of staurosporine or 3μM of cycloheximide. LDH released into the medium was measured along with blank, untreated cells (low control), LDH Positive Control, and lysed cells (high control)
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"