TBARS Assay Kit

Catalog No. ABIN1000279

Product Details

Target: TBARS

Application: Biochemical Assay (BCA)

Sample Type: Plasma, Saliva, Serum, Urine

Specificity: 0.1 μM

Characteristics: Sensitive and accurate. Linear detection range: colorimetric assay 1 - 30 µM, fluorometric assay 0.1 - 1.5 µM MDA.

Components: TBA Reagent: 25 mL. Standard: 50 µL 15 mM MDA. 10% Trichloroacetic acid (TCA): 25 mL.

Material not included: Pipetting devices, centrifuge tubes, centrifuge, clear flat-bottom uncoated 96-well plates, optical density or fluorescence plate readers, sonicator, water-bath or heat block.

Application Details

Application Notes: Direct Assays: serum, plasma, urine, saliva and other biological samples.
Drug Discovery/Pharmacology: effects of drugs on TBARS.

Protocol: Set up water bath or heat block and adjust the temperature to 100°C. Equilibrate all components to room temperature. Add 450 µL dH2O to the 15 mM Standard tube and mix (final1.5 mM MDA). Store unused Standard at -20°C for future use.
1. Standards. Mix 15 µL of the1.5 mM MDA with 735 µL dH2O (final 30 µMMDA). Dilute standard. Transfer 200 µL standards into labeled1.5-mL screw cap tubes. Samples: Transfer 200 µL of each sample into separate tubes.
2. Color reaction. To each of the standards and samples, add 200 µL TBA Reagent. Vortex tubes to mix and incubate at 100°C for 60 min. Cool down tubes to room temperature. Vortex and briefly centrifuge tubes.
3. Load 100 µL in duplicate from each tube to wells of a clear flat- bottom 96-well plate. Read OD at 535nm (525 to 545nm).

Calculation of Results:

Subtract blank OD or fluorescence intensity value (#4) from all standard and sample values. Plot the OD535nm or F against standard concentrations and determine the slope of the standard curve. Note: if calculated TBARS concentration is higher than 30 µM MDA (colorimetric assay) or 1.5µM MDA (fluorometric assay) , dilute sample in dH2O and repeat assay. Multiply the results by the dilution factor.

Restrictions: For Research Use only

Handling

Storage: -20 °C

References

Shanmugasundaram, Selvaraj: "Dietary lutein and fish oil interact to alter atherosclerotic lesions in a Japanese quail model of atherosclerosis." in: Journal of animal physiology and animal nutrition, Vol. 95, Issue 6, pp. 762-70, (2011) (PubMed).

Shanmugasundaram, Selvaraj: "Lutein supplementation alters inflammatory cytokine production and antioxidant status in F-line turkeys." in: Poultry science, Vol. 90, Issue 5, pp. 971-6, (2011) (PubMed).

Davison: "Innate immune responses to a single session of sprint interval training." in: Applied physiology, nutrition, and metabolism = Physiologie appliquée, nutrition et métabolisme, Vol. 36, Issue 3, pp. 395-404, (2011) (PubMed).

Verweij, van Ginhoven, Mitchell, Sluiter, van den Engel, Roest, Torabi, Ijzermans, Hoeijmakers, de Bruin: "Preoperative fasting protects mice against hepatic ischemia/reperfusion injury: mechanisms and effects on liver regeneration." in: Liver transplantation : official publication of the American Association for the Study of Liver Diseases and the International Liver Transplantation Society, Vol. 17, Issue 6, pp. 695-704, (2011) (PubMed).

Habib, Eisa, Arafat, Marie: "Pulmonary involvement in early rheumatoid arthritis patients." in: Clinical rheumatology, Vol. 30, Issue 2, pp. 217-21, (2011) (PubMed).

Target Details

Target: TBARS

Background: Quantitative determination of thiobarbituric acid reactive substances (TBARS) by colorimetric (535nm) or fluorimetric (560nm/585nm) methods.
Procedure: 80 min.

Oxidative attack of essential cell components by reactive oxygen species has been associated with several human diseases, such as atherosclerosis, cardiovascular diseases, diabetes, liver disorders, and inflammatory rheumatic diseases. Thiobarbituric Acid Reactive Substances (TBARS) are low-molecular-weight end products (mainly malondialdehyde, MDA) that are formed during the decomposition of lipid peroxidation products. Increased levels of TBARS have been demonstrated in these diseases. Simple, direct and accurate assays for TBARS find wide applications in research and drug discovery. This TBARS assay is based on the reaction of TBARS with thiobarbituric acid (TBA) to form a pink colored product. The color intensity at 535nm or fluorescence intensity at (ex/em = 560nm/585nm) is directly proportional to TBARS concentration in the sample.