Pierce™ LDH Cytotoxicity Assay Kit

Catalog number: 88953

Thermo Scientific™  Related applications: Cell Counting, Viability, & Cryopreservation | Cell Viability, Proliferation & Function

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Description

The Thermo Scientific Pierce LDH Cytotoxicity Assay Kit is a reliable colorimetric assay to quantitatively measure lactate dehydrogenase (LDH) released into the media from damaged cells as a biomarker for cellular cytotoxicity and cytolysis.

Features of the LDH Cytotoxicity Assay Kit:

Convenient—add-mix-read assay format for adherent and suspension cells
Colorimetric—quantitatively measures LDH release by formation of colored product
Robust—uses stable LDH enzyme activity as a cytotoxic marker
Flexible—ideal for high-throughput screening
Non-radioactive—safe alternative to 51Cr-release cytotoxicity assays

The Pierce LDH Cytotoxicity Assay Kit measures extracellular LDH in culture media using an enzymatic reaction that results in a red formazan product which can be measured spectrophotometrically. Lactate dehydrogenase (LDH) is a cytosolic enzyme that is is an indicator of cellular toxicity. The assay is ideal for high-throughput screening and provides a safe alternative to radioactive cytotoxicity assays.

Includes:
Kit contains substrate mix, assay buffer, 10X lysis buffer, stop solution, and LDH positive control

Requires:
Microplate reader capable of reading absorbance at 490nm and 680nm, flat-bottom clear 96-well plate compatible with spectrophotometer, multichannel pipette

Applications:
• Measure in vitro cytotoxicity mediated by chemicals, immune cells, or siRNA or microRNA
• Detect cytolysis in bioreactor or 3D tissue engineering applications

Lactate dehydrogenase (LDH) is a cytosolic enzyme present in many different types of cells. When the plasma membrane is damaged, LDH is released into cell culture media. The released LDH can be quantified by a coupled enzymatic reaction. First, LDH catalyzes the conversion of lactate to pyruvate via reduction of NAD+ to NADH. Second, diaphorase uses NADH to reduce a tetrazolium salt (INT) to a red formazan product. Therefore, the level of formazan formation is directly proportional to the amount of released LDH in the medium.

The assay is performed by transferring cell culture media from treated cells into a new microplate and adding the kit reagents. After incubation at room temperature for 30 minutes, reactions are stopped and LDH activity is determined by spectrophotometric absorbance at 490nm.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Compatible Cells: Mammalian Cells
For Use With (Equipment): Microplate Reader
Detection Method: Colorimetric
Format: 96-well plate, 384-well plate
Product Size: 200 reactions

Contents & storage

Kit Contents and Storage Information:

Substrate Mix, lyophilizate, store at -20°C protected from light
Assay Buffer, .6mL, store at -20°C protected from light
1X Lysis Buffer, 2.5mL, store at 4°C
Stop Solution, 12mL, store at 4°C
LDH Positive Control, 6µL, store at 4°C

Kit is shipped at ambient temperature.

Documents

Manuals & protocols