H2DCFDA (DCFH-DA)

製品コードS9687 バッチS968701

印刷

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

7 mg/mL (14.36 mM)

Water

Insoluble

体内（毎回新しく調製した物を用意してください）

Homogeneous suspension

CMC-NA

≥5mg/ml

Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.

* <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) は、細胞内の活性酸素種 (ROS) の産生を検出するために使用される細胞透過性プローブです。細胞内で脱エステル化され、酸化されると高蛍光性の2',7'-ジクロロフルオレセインに変化します。酸化代謝に応答する酸素反応性種の高感度かつ迅速な定量、食細胞における酸化産物を検出するためのマイクロプレートアッセイ、および定量的マルチウェル骨髄分化アッセイに使用されます。(Ex/Em=488/525 nm)
in vitro	1. H2DCFDA (DCFH-DA) is dissolved in DMSO to obtain a 10 mM stock solutions and further diluted before use. 2. Cells are incubated with 5 μM of this compound 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 it, 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) は、細胞内の活性酸素種 (ROS) の産生を検出するために使用される細胞透過性プローブです。細胞内で脱エステル化され、酸化されると高蛍光性の2',7'-ジクロロフルオレセインに変化します。酸化代謝に応答する酸素反応性種の高感度かつ迅速な定量、食細胞における酸化産物を検出するためのマイクロプレートアッセイ、および定量的マルチウェル骨髄分化アッセイに使用されます。(Ex/Em=488/525 nm)

in vitro

1. H2DCFDA (DCFH-DA) is dissolved in DMSO to obtain a 10 mM stock solutions and further diluted before use.
2. Cells are incubated with 5 μM of this compound 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 it, 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 H2DCFDA (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 this compound 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

https://pubmed.ncbi.nlm.nih.gov/26976009/
https://pubmed.ncbi.nlm.nih.gov/30372737/
https://pubmed.ncbi.nlm.nih.gov/26976009/

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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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