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Specifications
- Assay Type Detection Kit
- Sample Types Serum, Plasma, Tissue, Cell Extracts, Food Extracts, Urine, Buffers
- Sensitivity 0.36 µM
- Species Species Independent
- Assay Duration 1 Hour
- Samples/Plate 40 in duplicate
- Readout Colorimetric, 535 nm
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Standard Curve
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Description
Assay Principle:
The TBARS/MDA Universal Colorimetric Detection Kit quantitatively measures lipid peroxidation levels in serum, plasma, tissue, cell extracts, food extracts, urine, and buffers. The TBARS/MDA Universal Colorimetric Detection Kit is a Detection Kit with a run time of 60 minutes. Please read the complete kit insert for more information before performing this assay.
Use our provided MDA Standard to generate a standard curve for the assay. Pipette the standards or diluted samples into a transparent microtiter plate. Add TBA Substrate to each well tapping the plate to ensure sufficient mixing of reagents. Then incubate the mixture covered at 37°C shaking for 60 minutes. The color-generating reaction occurs between the TBA substrate and the MDA within the sample or standard.
After the 60-minute incubation, use a plate reader to detect and record the signal generated at 535nm. Use the intensity and the standard curve to calculate the MDA concentration in the samples.
Background:
Lipid peroxidation is a well-established mechanism of cellular injury in plants and animals and is used as an indicator of oxidative stress in cells and tissues. Lipid peroxidation products derived from polyunsaturated fatty acids are stable and decompose to form a diverse mixture of compounds, including MDA.
MDA is conveniently measured by the reaction of thiobarbituric acid in an acid environment according to the following reaction.
The MDA-TBA adduct formed in this reaction is pink-colored and can be read at ƛ= 532 nm. Measurement between 530–545 nm is acceptable.
In addition to MDA, these oxidative mechanisms can form other reactive aldehydes that react with TBA to generate color, including 2-alkenals and 2,4-alkedienals. The combined total of MDA plus other reactive substances is TBARS for Thiobarbituric Acid Reactive Substances. The TBARS assay is modified to evaluate several samples, including mammalian tissues, serum, plasma and urine, and food samples. There is some ambiguity surrounding the use of TBARS in different sample types under different oxidative stress because of the reactivity of acidified TBA toward reactive aldehydes. However, the assay is used extensively to determine lipid peroxidation. In general, lipids with greater unsaturation will yield higher TBARS values.