H2DCFDA (DCFH-DA)

製品コードS9687 バッチS968702

化学情報

	Synonyms	2',7'-Dichlorodihydrofluorescein diacetate	Storage (From the date of receipt)	3 years -20°C(in the dark) powder
	化学式	C₂₄H₁₆Cl₂O₇
	分子量	487.29	CAS No.	4091-99-0
Solubility (25°C)*	体外	DMSO	97 mg/mL (199.06 mM)
		Ethanol	15 mg/mL (30.78 mM)
		Water	Insoluble
* <1 mg/ml means slightly soluble or insoluble. * Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

溶剤液（一定の濃度）を調合する

生物活性

製品説明	H2DCFDA (DCFH-DA) is a cell-permeable probe used to detect intracellular reactive oxygen species (ROS) production. It is de-esterified intracellularly and turns to highly fluorescent 2′,7′-dichlorofluorescein upon oxidation. It is used in sensitive and rapid quantitation of oxygen-reactive species in response to oxidative metabolism; microplate assay for detecting oxidative products in phagocytic cells, and quantitative multiwell myeloid differentiation assay.(Ex/Em=488/525 nm)
in vitro	1. H2DCFDA is dissolved in DMSO to obtain a 10 mM stock solutions and further diluted before use. 2. Cells are incubated with 5 μM H2DCFDA 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. 3. Along with the H2DCFDA probe, if indicated, use ROS-insensitive modification of the fluorescein dye DCFDA as a positive control. The staining procedure is the same as for the H2DCFDA。 Emission (Em)=517 Excitation (Ex)=492 517

製品説明

H2DCFDA (DCFH-DA) is a cell-permeable probe used to detect intracellular reactive oxygen species (ROS) production. It is de-esterified intracellularly and turns to highly fluorescent 2′,7′-dichlorofluorescein upon oxidation. It is used in sensitive and rapid quantitation of oxygen-reactive species in response to oxidative metabolism; microplate assay for detecting oxidative products in phagocytic cells, and quantitative multiwell myeloid differentiation assay.(Ex/Em=488/525 nm)

in vitro

1. H2DCFDA is dissolved in DMSO to obtain a 10 mM stock solutions and further diluted before use.
2. Cells are incubated with 5 μM H2DCFDA 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.
3. Along with the H2DCFDA probe, if indicated, use ROS-insensitive modification of the fluorescein dye DCFDA as a positive control. The staining procedure is the same as for the H2DCFDA。
Emission (Em)=517 Excitation (Ex)=492 517

プロトコル(参考用のみ)

細胞アッセイ	細胞株	hCECs
	濃度	10 μM
	反応時間	30 min
	実験の流れ	Intracellular ROS levels in hCECs are measured by incubating the cells with the redox-sensitive fluorescent dye 2'7'-dichlorofluorescein diacetate (DCFH-DA) (10 μM) at 37℃ for 30 minutes in the dark. The cells are then detached from the culture wells using 0.25% trypsin-EDTA and washed twice using ice-cold PBS. Flow cytometry measurements are performed three times for each treatment. Mean fluorescence intensity is quantified using software. Cells that had been incubated without DCFH-DA are used as negative control. The distribution of fluorescent dichlorofluorescein (DCF) on the cell monolayer is visualized and photographed using a fluorescence microscope.

参考

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長期の保管のために-20°Cの下で製品を保ってください。

人間や獣医の診断であるか治療的な使用のためにでない。

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