H2DCFDA

Biological Activity

H2DCFDA is a cell-permeable probe used to detect intracellular reactive oxygen species (ROS).

ROS Measurements

For the detection of intracellular ROS level ,ROS-sensitive probe H2DCFDA is used. Adherent cells (ESCs, difESCs, eMSCs, HeLa, U118) are incubated with 5 µM staining solution in PBS in the dark for 30 min at 37°C, then harvested with 0.05% trypsin-EDTA solution, suspended in a fresh medium, and immediately analyzed with flow cytometer. Lymphocytes, both control and PHA-activated, are resuspended in PBS, incubated with 5 µM of H2DCFDA in the dark for 30 min at 37°C, and immediately analyzed. Along with the H2DCFDA probe, if indicated, ROS-insensitive modification of the fluorescent dye DCFDA is used. The staining procedure is the same as for the H2DCFDA.

Product Citations

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  Ren Fail. 2021 Dec;43(1):900-910.

  MMP-9-mediated regulation of hypoxia-reperfusion injury-related neutrophil inflammation in an in vitro proximal tubular cell model
  H2DCFDA purchased from AbMole

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  Mediators Inflamm. 2020 Jun 6;2020:4321912.

  CD137 Signaling Promotes Endothelial Apoptosis by Inhibiting Nrf2 Pathway, and Upregulating NF- κ B Pathway
  H2DCFDA purchased from AbMole

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  Comp Biochem Physiol C Toxicol Pharmacol. 2021 Apr;242:108938.

  Clove essential oil confers antioxidant activity and lifespan extension in C. elegans via the DAF-16/FOXO transcription factor
  H2DCFDA purchased from AbMole

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  Anat Rec (Hoboken). 2020 Jan 21.

  Resibufogenin inhibited colorectal cancer cell growth and tumorigenesis through triggering ferroptosis and ROS production mediated by GPX4 inactivation
  H2DCFDA purchased from AbMole

Customer Product Validations & Biological Datas

	Source	Comp Biochem Physiol C Toxicol Pharmacol (2021 Apr), Figure 3, H2DCFDA (Abmole Bioscience)
	Method	measure the ROS levels
	Cell Lines	Each C. elegans strain
	Concentrations	50 μM
	Incubation Time	1 h
	Results	The results showed that CEO exposure made no significant difference to the GSH amount in each worm, however, the lipid peroxidation degree decreased due to the reduced ROS levels.

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

Species	Mouse	Rat	Rabbit	Guinea pig	Hamster	Dog
Weight (kg)	0.02	0.15	1.8	0.4	0.08	10
Body Surface Area (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km factor	3	6	12	8	5	20

Animal A (mg/kg) = Animal B (mg/kg) multiplied by	Animal B Km
	Animal A Km

For example, to modify the dose of Compound A used for a mouse (20 mg/kg) to a dose based on the BSA for a rat, multiply 20 mg/kg by the K_m factor for a mouse and then divide by the K_m factor for a rat. This calculation results in a rat equivalent dose for Compound A of 10 mg/kg.

Chemical Information

References

Redox environment in stem and differentiated cells: A quantitative approach.
Lyublinskaya OG, et al. Redox Biol. 2017 Aug;12:758-769. PMID: 28426982.

Reactive Oxygen Species in Planarian Regeneration: An Upstream Necessity for Correct Patterning and Brain Formation
Nicky Pirotte, et al. Oxid Med Cell Longev. 2015;2015:392476. PMID: 26180588.

Sustained production of ROS triggers compensatory proliferation and is required for regeneration to proceed
Carole Gauron, et al. Sci Rep. 2013;3:2084. PMID: 23803955.