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Pierce LDH Cytotoxicity Assay Kit


A colorimetric assay for measuring cellular cytotoxicity.

Pierce LDH Cytotoxicity Assay Kit

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

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.

Highlights:

  • 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

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

Product Details:

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.

Fluorescence and absorbance spectra of alamarBlue Cell Viability Reagent
Metabolic and chemical basis of the LDH cytotoxicity assay.

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.

Protocol summary for alamarBlue Cell Viability Reagent.
Protocol summary for Thermo Scientific Pierce LDH Cytotoxicity Assay Kit.

 

Cell number optimization  for alamarBlue Cell Viability Reagent.
Determination of Actinomycin D-induced cytotoxicity in CHO-K1 cells using the Thermo Scientific Pierce LDH Cytotoxicity Assay. CHO-K1 cells were plated in a 96-well plate (10,000 cells per well) in RPMI1640 medium supplemented with 10% fetal bovine serum and incubated overnight at 37°C in 5% CO2. Cells were then treated with different concentrations of the cytotoxic agent, Actinomycin D, for 18 hours. Actinomycin-induced cytotoxicity was measured using the Pierce LDH Cytotoxicity Assay Kit (red; left y-axis). Cell viability was measured in replicate wells using the alamarBlue Cell Viability Reagent (Part No. 88951, 88952)(blue; right y-axis).

General References:

  1. Decker, T. and Lohmann-Matthes, M.L. (1988). A quick and simple method for the quantitation of lactate dehydrogenase release in measurements of cellular cytotoxicity and tumor necrosis factor (TNF) activity. J. Immunol. Meth. 115:61-9.
  2. Korzeniewski, C. and Callewaert, D.M. (1983). An enzyme-release assay for natural cytotoxicity. J. Immunol. Meth. 64:313-20.

Related Products:

alamarBlue Cell Viability Assay Reagent
Cellular Imaging and Analysis Reagents (formerly Cellomics Products)


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
88953 LDH Cytotoxicity Assay Kit
Formulation: Five-component kit
Sufficient For: 200 reactions in 96-well plates or 800 reactions in 384-well plates

Kit Contents:
Substrate Mix, lyophilizate
Assay Buffer, 0.6mL
10X Lysis Buffer, 2.5mL
Stop Solution, 12mL
LDH Positive Control, 6μL
200-rxn kit Product Instructions for product #88953 LDH Cytotoxicity Assay Kit MSDS for product #88953 LDH Cytotoxicity Assay Kit Certificate of Analysis for product #88953 LDH Cytotoxicity Assay Kit $109.00

Add to cart
88954 LDH Cytotoxicity Assay Kit
Formulation: Five-component kit
Sufficient For: 1000 reactions in 96-well plates or 4000 reactions in 384-well plates

Kit Contents:
Substrate Mix, 5 x lyophilizate
Assay Buffer, 5 x 0.6mL
10X Lysis Buffer, 12mL
Stop Solution, 60mL
LDH Positive Control, 30┬ÁL
1000-rxn kit Product Instructions for product #88954 LDH Cytotoxicity Assay Kit MSDS for product #88954 LDH Cytotoxicity Assay Kit Certificate of Analysis for product #88954 LDH Cytotoxicity Assay Kit $297.00

Add to cart

Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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