How can I prepare the standard curve of Malondialdehyde (MDA) especially with Tetraethoxypropane (TEP) for TBARS calculation in UV spectrophotometer. Can anyone suggest how should I prepare the standard solution of TEP and in what range ?
Dear Dr Carpenter,
May lab protocol was as follows in my recent submitted work:
2.9 Estimation of lipid peroxidation (malondialdehyde (MDA) assay)
The magnitude of lipid peroxidation was determined by measuring MDA which is thiobarbituric acid reactive substance (TBARS). To precipitate the proteins, TCA (30%; 0.5 ml) was added to tissue extract (0.5 ml), vortexed for 30 seconds, and finally centrifuged at 3,000 rpm for 5 minutes. Thiobarbitoric acid (TBA; 1%; 500 µl) solution and 500 µl of distilled water were added to the supernatant and the resulting mixture heated for 1 hour at 98 °C, then cooled to room temperature and kept in ice for 5 minutes. At last, the absorbance of pink mixture recorded at 532 nm. Standard graph was plotted using 1,1,3,3-tetraethoxy propane (TEP) to estimate MDA values [30].
[30] Ohkawa, H., Ohishi, N., Yagi, K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Analytical Biochemistry. 1979,95:351-8.
Thank you Dr. Karimi for your reply and suggestion. I got the way of MDA measurement with the above procedure suggested by you. Now, to prepare the standard curve of TEP, should I just use TEP (as MDA source) instead of my sample and keeping the whole procedure same ? Is this the way to prepare the standard of TEP or something else ?
Standard method for Malondialdehyde (MDA) measurement using Tetramethoxypropane for TBARS calculation in UV spectrophotometer ?
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