H2DCFDA试剂
H2DCFDA (DCFH-DA) 是一种可渗透细胞的，用于检测细胞内活性氧 (ROS) 的探针 (Ex/Em=488/525 nm)。
H2DCFDA试剂生物活性
H2DCFDA (DCFH-DA) is a cell-permeable probe used to detect intracellular reactive oxygen species (ROS) (Ex/Em=488/525 nm)
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| 运输条件 | Room temperature in continental US; may vary elsewhere. |
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| 储存方式 | -20°C, protect from light |
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| 溶解性数据 | In Vitro: DMSO : 125 mg/mL (256.52 mM; Need ultrasonic) Ethanol : 20 mg/mL (41.04 mM; Need ultrasonic) 配制储备液 | 1 mM | 2.0522 mL | 10.2608 mL | 20.5217 mL | | 5 mM | 0.4104 mL | 2.0522 mL | 4.1043 mL | | 10 mM | 0.2052 mL | 1.0261 mL | 2.0522 mL |
*请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month (protect from light)。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 In Vivo: 请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂： ——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶 1.
请依序添加每种溶剂： 10% DMSO 40% PEG300 5% Tween-80 45% saline Solubility: 2.08 mg/mL (4.27 mM); Suspended solution; Need ultrasonic
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请依序添加每种溶剂： 10% DMSO 90% (20% SBE-β-CD in saline) Solubility: ≥ 2.08 mg/mL (4.27 mM); Clear solution
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请依序添加每种溶剂： 10% DMSO 90% corn oil Solubility: ≥ 2.08 mg/mL (4.27 mM); Clear solution
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| 染色示例 | Description: H2DCFDA (DCFH-DA) is a ROS production probe with green fluorescence.
Method: For cell staining.
1. Stain cells with H2DCFDA.
2. Capture staining cells by spinning-disk confocal microscopy (every 5 min for 6 h). Description: H2DCFDA (DCFH-DA) is a ROS production probe with green fluorescence.
Method: For cell staining.
1. Remove the cell culture medium and replaced by fresh medium containing H2DCFDA and incubated with cultured cells for 30 min.
2. Wash cells with PBS for three times.
3. Use a confocal laser scanning microscopy (FV1000, Olympus Company, Japan) to detect the intracellular ROS generation. Description: H2DCFDA (DCFH-DA) is a ROS production probe with green fluorescence.
Method: For cell staining.
1. Testing cells are seeded into a 6-well microplate at a density of 105/well and cultured overnight at 37°C in 5% CO2. Fresh medium with 50 μg/mL of different materials is added to each well and cultured for 16 h.
2. Wash cells with PBS and stain cells with H2DCFDA (20 μM; 1 h; 37°C; dark).
3. Cells are trypsinized, washed, resuspended in PBS (0.2 mL), analyzed by a flow cytometer and use a confocal laser scanning microscopy (FV1000, Olympus, Tokyo, Japan) for image. Description: H2DCFDA (DCFH-DA) is a ROS production probe with green fluorescence.
Method: For cell staining.
1. Wash cultured cells with PBS for three times.
2. Treat testing cells with H2DCFDA in culture medium for 0.5 h in a cell incubator.
3. Use a confocal laser scanning microscopy (Olympus, FV3000) for image. Cells are analyzed on a BD LSRI Fortessa flow cytometer analyzer to quantify results. Description: H2DCFDA (DCFH-DA) is a ROS production probe with green fluorescence.
Method: For cell staining.
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