Cardiovascular and Oxidative Stress Assay Kit

TBARS Cuvette Assay Kit

$490.00

The TBARS Cuvette Assay kit is intended for the quantitative determination of 2-Thiobarbituric Acid Reactive Substances in serum, plasma, or urine. The TBARS Cuvette Assay Kit is for research use only and not to be used in diagnostic procedures.

SKU: TBA39-K01 Categories: , ,

TBARS Cuvette Assay Kit

The TBARS Cuvette Assay Kit is For Research Use Only

Size: 100 Tests
Sensitivity: 0.5 µM
Standard Range: 0.5 – 20 µM
Incubation Time: 45 minutes
Sample Type: Serum, Plasma, Urine
Sample Size: 400 µL

Assay Background

2-ThioBarbituric Acid Reactive Substances (TBARS) are naturally present in biological specimens and include lipid hydroperoxides and aldehydes which increase in concentration as a response to oxidative stress. TBARS assay values are usually reported in malonaldehyde (malondialdehyde, MDA) equivalents, a compound that results from the decomposition of polyunsaturated fatty acid lipid peroxides. The TBARS assay is a well-recognized, established method for quantifying these lipid peroxides, although it has been criticized for its reactivity towards other compounds other than MDA. This TBARS Cuvette Assay offers the researcher a straightforward, reproducible and consistent method for analyzing urine for lipid peroxidation products.

SAMPLE PREPARATION
When working with plasma or serum, the sample should be deproteinated with an acid. Centrifuge and use the supernatant to perform the assay. This solution may appear cloudy after the reaction, and can be clarified by passing through a 0.2 µ syringe filter. When working with urine, colored compounds contribute to the signal measured at 532 nm. This interference can be removed by running a sample blank with each sample.
Urine:
1. Urine samples can be used directly and should be assayed immediately. If the assay is to be performed on a different day, the sample should be stored at -70°C.

Plasma and Serum
1. Collect blood samples and process immediately per the collection tube instructions.
2. Prepare a saturated solution of ammonium sulfate.
3. Add 200 µL of saturated ammonium sulfate to 1.0 mL of serum or plasma in a test tube or microcentrifuge tube.
4. Add 75 mg TCA (trichloroacetic acid) to each sample and vortex. A cloudy precipitate should form.
5. Centrifuge the tubes and transfer the supernatant to a clean tube. Plasma and serum samples can be run without dilution. Samples are now ready to assay.

Related Products

TBARS ELISA Assay Kit
TBARS Fluorometric Assay Kit
TBARS Assay for Food and Beverages

Additional Information

Assay Principle

This TBARS Cuvette Assay Kit is based on the reaction of a chromogenic reagent, 2-thiobarbituric acid, with MDA at 25°C. One molecule of MDA reacts with 2 molecules of 2-thiobarbituric acid via a Knoevenagel-type condensation to yield a chromophore with absorbance maximum at 532 nm.

Assay Procedure

Free MDA

Preparation of Standards and Samples: Add each of the following reagents into glass test tubes and mix well.

  • Standards: 0.3 mL of standard and 0.3 mL of Indicator Solution.
  • Samples: 0.3 mL of sample and 0.3mL of Indicator Solution.
  • Blanks: 0.3 mL of sample and 0.3mL of Acid Reagent

After the standards, samples and blanks have been mixed; allow them to react for 45 minutes at room temperature.

For Colorimetric reading, transfer 0.5 mL of each solution to cuvettes and read at 532 nm. The pink color is stable for several hours at room temperature.

For Fluorometric reading, transfer 0.5 mL of each solution to cuvettes and excite the sample at 532 nm and read at 585nm.

Total MDA

Prepare samples and standards exactly as above, but heat sample at 65˚C for 45 minutes, then follow step 3 as above.

Documents

Product Documents

Publications

Citations

Lačná, Júlia, František Foret, and Petr Kubáň. “Sensitive determination of malondialdehyde in exhaled breath condensate and biological fluids by capillary electrophoresis with laser induced fluorescence detection.” Talanta169 (2017): 85-90. Web.

Product Citations