Cardiovascular and Oxidative Stress Assay Kit

Lipid Peroxidase Assay Kit

$435.00

The Lipid Peroxidase Assay kit is for the quantitative determination of either malondialdehyde (MDA) alone (in hydrochloric acid) or MDA in combination with 4-hydroxyalkenals (HAE) (in methanesulfonic acid.) in biological fluids by a chromogenic reagent microplate assay.

SKU: LIP39-K01 Categories: , ,

Lipid Peroxidase Assay Kit

The Lipid Peroxidase Assay Kit is For Research Use Only

Size: 1×96 wells
Sensitivity: 0.1 nmol/mL
Dynamic Range: 0.5 – 15 µM
Incubation Time: 3 hours
Sample Type: Biological Fluids
Sample Size: 140 µl
Alternative Name: LPO

Assay Background

Lipid peroxidation is a well-established mechanism of cellular injury in both plants and animals, and is used as an indicator of oxidative stress in cells and tissues. Lipid peroxides are unstable and decompose to form a complex series of compounds including reactive carbonyl compounds. Polyunsaturated fatty acid peroxides generate malondialdehyde (MDA) and 4-hydroxyalkenals (HAE) upon decomposition. The measurement of MDA and HAE has been used as an indicator of lipid peroxidation (1).

SAMPLE PREPARATION

Sample Stability:
Unless assayed immediately, samples should be frozen at -70°C to prevent loss of MDA (3,4) and prevent new sample oxidation. Samples should not be stored at -20°C. Once thawed from -70°C storage for assay, the sample should not be refrozen. Samples should be protected from light to avoid photoxidation.

Oxidation Prevention:
We recommend adding BHT at a final concentration of 5 mM in the buffer prior to homogenization of tissue or cells. If no antioxidant is added, new lipid peroxidation can occur during homogenization and biased values will result (2).

Plasma or Serum:
The amount of free MDA in normal plasma or serum is at or below the limit of detection of this assay.

Cell Culture:
Cells cultured in serum containing medium should be washed several times to remove serum components prior to homogenization. Since MDA exists as the water-soluble enolate anion at physiological pH, much of the MDA generated from lipid peroxidation in cell culture may be in the culture medium.

Related Products

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Creatinine Microplate Assay Kit
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Additional Information

Assay Principle

This Lipid Peroxidation Assay is based on the reaction of a chromogenic reagent, N-methyl-2-phenylindole (R1), with MDA and HAE at 45°C. One molecule of MDA reacts with 2 molecules of Reagent R1 to yield a stable chromophore with maximal absorbance at 586 nm.

Assay Procedure

  1. Add 140 µL of Standards or Samples to a microcentrifuge tube.
  2. Add 455 µL of diluted Reagent R1 to each tube and vortex.
  3. Add 105 µL 37% HCl (12 N HCl) to each tube and mix well.
  4. Incubate at 45°C for 60 minutes.
  5. Centrifuge samples at 15,000 x g for 10 minutes to obtain a clear supernatant.
  6. Transfer the 3 x 150 µL of the supernatant to the microplate and read at 586 nm.

See Scheme I for a sample plate layout. The color is stable for at least an hour at room temperature, or 2 hours at 4°C when stored in the dark.

Package Inserts

Package Inserts

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

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