What is best protocol for estimation of proline, MDA content Catalase and SOD for plant leaves ?
Reagents:
- 1% solution of phosphoric acid;
- 0.8% solution of thiobarbituric acid (TBA);
- n-butanol.
Conducting research.
To 0.5 ml (supernatant of fresh plant material homogenate) add 3.0 ml of 1% phosphoric acid solution and 1.0 ml of 0.8% TBA solution. The tubes are placed in a boiling water bath for 1 hour. Then the tubes are cooled in cold water, 4.0 ml of n-butanol are added, thoroughly mixed and centrifuged for 10 min at 3000 rpm (at a temperature of 15 °C). Measure the optical density of the upper phase at a wavelength of 532 nm against n-butanol. The calculation of the content of TBA-dependent products is carried out taking into account the molar extinction coefficient of MDA, equal to 1.56 . 105 M-1cm-1, according to the formula:
,
where: V is the volume of n-butanol extract;
E - sample extinction;
a - a sample of fabric;
N is the dilution index.
The MDA content is expressed in µmol/g sample.
Best wishes,
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