TBARS ELISA Assay Kit

$305.00

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

SKU: TBR39-K01 Categories: , ,

TBARS ELISA Assay Kit

The TBARS ELISA Assay Kit is For Research Use Only

Size: 1 x 96 wells
Sensitivity: 0.5 µM
Standard Range: 0.5 – 20 µM
Incubation Time: 30 minutes
Sample Type: Serum, Plasma, Urine
Sample Size: 100 µl

Product Developed and Manufactured in the USA


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.1,2 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.3 This kit offers the researcher a straightforward, reproducible and consistent method for analyzing urine for lipid peroxidation products.


STORAGE
1. The reagents of the TBARS ELISA are stable until the indicated kit expiration date if handled and stored properly.
2. When not in use, store the TBARS ELISA Assay Kit at 4°C for up to one year.
3. MDA standards should be used within 24 hours of preparation.
4. The Indicator Solution (combined 2-TBA and Acid Solution) can be stored at 4˚C forone week.


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Additional Information

Assay Principle


This TBARS ELISA 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

Free MDA

  1. Preparation of Standards and Samples:  Add each of the following reagents into microcentrifuge tubes and mix well.
    • Standards:     100 μL of standard and 300 μL of Indicator Solution.
    • Samples:     100 μL of sample and 300 μL of Indicator Solution.
    • Blanks:     100 μL of sample and 300 μL of Acid Reagent.
  2. After the standards, samples and blanks have been mixed; allow them to react for 20 minutes at room temperature.
  3. Transfer 150 µL of each solution to the microplate and read at 540 nm. Please note that the optimum absorbance wavelength for this assay is 532 nm. The pink color is stable for several hours at room temperature.

Total MDA
Prepare samples and standards exactly as above, but heat sample at 65°C for 30 minutes, then follow step 3 as above.

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References


  1. Armstrong, D.; Browne, R. (1994) Free Rad. Diag. Med. 366; 43-58.
  2. Botsoglou, N.A., et al.; (1994) J. Agric. Food Chem. 42; 1931-1937.
  3. Yagi, K. (1998) Free Rad. Antiox. Prot., 108; 101-106.
  4. Moore, K., Roberts, L.J., (1998) Free Rad. Res. 28; 659-671.
  5. Liu, J., et al.; (1997) Analyt. Biochem. 245;161-166.
  6. Carbonneau, M.A., et al.; (1991) Clin. Chem. 37; 1423-1429.
  7. Mattson, J.P., et al.; (2002) Pathophysiology 8; 215-221.
  8. Calvo, J.R., et al.; (2001) J. Cell Biochem. 80; 461-70.
  9. Janciauskiene, S. and Bo Ahren; (2000) Biochem. Biophys. Res. Com. 267; 619-625.
  10. Montilla-Lopez, P., et al.; (2002) European J. Pharmacology 451; 237-243.

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