TBARS Malondialdehyde Oxidative Stress Assay

$282.75

Product #: 1020

Specimen Required: 100 µl or 50 µl

Assay Range: 0.5- 80 µm

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Description

TBARS Malondialdehyde Oxidative Stress Assay

Our Exocell™ TBARS Malondialdehyde Oxidative Stress assay measures thiobarbituric acid reactive substances in biological specimens.

Reactive oxygen species are a consequence of oxidative metabolism, but are balanced by reducing capacity under normal conditions.  However, through injury or disease this balance may be lost and free radical mediated damage can occur.  Lipid peroxidation is an example of this damage.

Lipid peroxidation can affect biological membranes by altering structure, permeability and function.  It is implicated in a number of disease states:  premature birth disorders, diabetes, adult respiratory distress syndrome, shock, Parkinson’s disease, Alzheimer’s disease, chronic inflammatory conditions and ischemia-reperfusion mediated injury to kidney, heart, brain and intestine (1).

Upon the metabolic breakdown of the altered lipids is the formation of “secondary messengers” including malondialdehyde (MDA), 4-hydroxynonenal and others.  Measurement of MDA allows an assessment of oxidative stress.

The TBARS Malondialdehyde Oxidative Stress Assay depends on the reaction of thiobarbituric acid and MDA under acidic conditions to form a colored (and fluorescent) adduct (2).  The assay uses a substrate that is converted to MDA as a standard, and results are reported in MDA equivalents.  The response of the assay is linear over the range 0.5-80 µm .

Our kit is designed to allow a ready assessment of TBARS MDA equivalents in biological samples using either fluorometric or spectrophotometric methods.  The assay can be completed in volumes that allow assessments to be completed in standard cuvettes or in microplates.

  1. TPA Devasagayam et al.   Methods for estimating lipid peroxidation: an analysis of merits and demerits.  Indian J Biochem Biophys 40: 300-308.
  2. K Yagi.   A simple fluorometric assay for lipid peroxide in blood plasma.  Biochem Med 15: 212-216.

Kit Contents:

  1. TBA Stock Solution
  2.  MDA Standard: 300 µm
  3. TBARS Assay Diluent: 10X PBS must be diluted with water before use
  4. TBARS Reaction Buffer: 50% Acetic Acid. 2.5 mL of TBA stock is added to this buffer to make TBA working reagent.
  5. Microtiter plate that may be used with a plate reader in either fluorometric or in a spectrophotometric modes.

Materials required, but not supplied in the kit:

  1. Reaction tubes: for 500 µl reaction volumes, 1.7 mL microfuge tubes work well.  For 2.5 mL reaction volumes, 16 x 120 mm screw-cap glass tubes work well.
  2. Water bath or heat block set at 75o C
  3. Adjustable pipets
  4. Disposable pipets
  5. Graduated cylinder
  6. Purified water
  7. Disposable gloves
  8. Assay detection device:
    1. Plate reader: fluorometry:  ex: 532 nm/em 553 nm
    2. Plate Reader Spectrophotometry: OD 532 nm
    3. Fluorometer
    4. UV-Vis Spectrophotometer

Additional information

Weight 2 lbs
Dimensions 11 × 10 × 15 in

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