ROS assay cell type - PLHC-1, SK-HEP-1, Hep3b, HepG2 human hepatocellular carcinoma

ROS has a very short half-lives in biological environment as they are influenced by exposure to ambient oxygen. As it is highly reactive and hard to measure care should be taken to ensure the stability of the sample during isolation, preparation, storage, and analysis.

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Found 7 matching solutions for this experiment

Upstream tips
- Cells should be healthy and not overcrowded since results of the experiments will depend significantly on the cells’ condition.
Protocol tips
- The cells were stained using the two‐color ROS Detection Kit.

-Cells should be pre-treated with the ROS Inhibitor at least 30 minutes prior to induction.
Protocol tips
- 1X DCFH-DA/media solution contains 5% methanol. For cells that are sensitive to methanol,
we recommend instead preparing a 0.1X (100 µM) solution of DCFH-DA in cell culture media.
Downstream tips
- Due to light-induced auto-oxidation, DCFH-DA solutions at any concentration must be
protected from light.
Upstream tips
- 2 × 10^5 cells/well were seeded.

- Cells were either treated with or without diosgenin.
Protocol tips
- Cells were incubated with 5 μM carboxy-H2DCFDA for 15 min at 37°C.

- Cells were washed with PBS twice, trypsinized, and resuspended in OptiMem I medium.
Upstream tips
- Cells were cultured and treated with phorbol myristate acetate (PMA) and ionomycin and incubated for 30 min.

- 5 × 10^5 were used for as ROS assay.
Protocol tips
- 10 µM final concentration of Carboxy-H2DCFDA was used incubated for 30 min.
Upstream tips
- 2.5 × 104 cells were seeded for an assay
Downstream tips
- The fluorescence intensity was measured using a fluorescence plate reader at Ex/Em. = 488/525 nm.
Downstream tips
- Due to light-induced auto-oxidation, the stock DCF-DiOxyQ solution and all subsequent
DCF-DiOxy and DCFH solutions must be protected from light.
Upstream tips
- 5 × 10^4 cells were seeded for the assay.

Protocol tips
- As a negative control cells were treated with 15 µg/mL NaB
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