Dihydroethidium

DNA quantification Human - SH-SY5Y

Experiment
DNA quantification Human - SH-SY5Y
Product
Dihydroethidium from Sigma-Aldrich
Manufacturer
Sigma-Aldrich

Protocol tips

Upstream tips
This product should be kept under argon. It is both light and air sensitive. It will oxidize to ethidium on storage unless kept under argon.

Publication protocol

Fluorometric measurement of superoxide anion production
Superoxide anion production was detected using the dihydroethidium (DHE) probe. 5 × 105 cells/well were seeded on 6-well tissue culture plates and allowed to adhere for 24 h in a 5% CO2 incubator at 37 0C. Thereafter, cells were treated with Oxa and Mel in their respective medium. Further, cells were stained or incubated with DHE solution (2 μg/ml) for 20 min. DHE dye incorporated into the cells and subsequently oxidized to the fluorescent ethidium cation by superoxide anion, allowing the cation to bind nuclear DNA with an extensive enhancement of fluorescence at 530 nm when excited at 485 nm [23]. The fluorescence signal was detected as proportional to superoxide production. The DHE fluorescence integrated density was analyzed with Image J software (1.50 version, NIH, USA). The 3–4 images were captured using NIS elements software provided by the Nikon Ts Eclipse inverted microscope at 20x magnification. Maximum projections were generated from focus planes and images were displayed with equal pixel intensity. Results were expressed as percentage of DHE stained positive cells.

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Discussion

Discussion

3 years ago

Author: Dorota Ostrowska Poland

About nuclease-free water in DNA quantification

Hi everyone! I have a question about DNA quantification. Do I need to use nuclease-free water for DNA quantification and are there any repercussions if I don’t use any?

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing DNA quantification Human - SH-SY5Y using Dihydroethidium from Sigma-Aldrich.

Paper title
Melatonin pre-treatment mitigates SHSY-5Y cells against oxaliplatin induced mitochondrial stress and apoptotic cell death
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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for Dihydroethidium below.

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