H2DCFDA 活性氧(ROS)探针

生物活性

H2DCFDA是可渗透细胞，用于检测细胞内活性氧 (ROS) 的探针 (Ex/Em=488/525 nm)。H2DCFDA可以用于细菌活性氧（ROS）产生的测定。

ROS测量

将H2DCFDA溶解在DMSO中以获得10mM储备溶液，并在使用前进一步稀释。

为了检测细胞内ROS水平，使用ROS敏感性探针H2DCFDA。将贴壁细胞（ESC，difESC，eMSC，HeLa，U118）与PBS中的5μM染色溶液在37℃下在黑暗中孵育30分钟，然后用0.05%胰蛋白酶-EDTA溶液收获，悬浮在新鲜培养基中，并且用流式细胞仪立即分析。将对照和PHA活化的淋巴细胞重悬浮于PBS中，与5μM H2DCFDA在黑暗中于37℃温育30分钟，并立即分析。

使用AbMole产品发表的文献

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产品使用成果展示

	数据来源	Comp Biochem Physiol C Toxicol Pharmacol (2021 Apr), Figure 3, H2DCFDA (Abmole Bioscience)
	方法	measure the ROS levels
	细胞系/动物模型	Each C. elegans strain
	浓度	50 μM
	处理时间	1 h
	实验结果	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.

化学性质

分子量	487.29
分子式	C24H16Cl2O7
CAS号	4091-99-0
溶解性（25°C）	DMSO: ≥ 90 mg/mL
储存条件	-20°C, protect from light
运输方式	冰袋运输，根据产品的不同，可能会有相应调整。

储备液配制

*下述溶液配置方法仅为基于分子量计算出的理论值。不同产品在配置溶液前，需考虑其在不同溶剂中的溶解度限制。

Concentration / Solvent Volume / Mass	1 mg	5 mg	10 mg
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

不同实验动物依据体表面积的等效剂量转换表（参考来源于公开文献）

	小鼠	大鼠	兔	豚鼠	仓鼠	狗
重量 (kg)	0.02	0.15	1.8	0.4	0.08	10
体表面积 (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km 系数	3	6	12	8	5	20

动物 A (mg/kg) = 动物 B (mg/kg) ×	动物 B的Km系数
	动物 A的Km系数

例如，依据体表面积折算法，将化合物用于小鼠的剂量20 mg/kg 换算成大鼠的剂量，需要将20 mg/kg 乘以小鼠的K_m系数（3），再除以大鼠的K_m系数（6），得到化合物用于大鼠的等效剂量为10 mg/kg。

参考文献

[1] Karyne Rangel, et al. Microorganisms. Detrimental Effect of Ozone on Pathogenic Bacteria

[2] Shijing Liao, et al. Int J Nanomedicine. Antibacterial activity and mechanism of silver nanoparticles against multidrug-resistant Pseudomonas aeruginosa

[3] Lyublinskaya OG, et al. Redox Biol. Redox environment in stem and differentiated cells: A quantitative approach.

[4] Nicky Pirotte, et al. Oxid Med Cell Longev. Reactive Oxygen Species in Planarian Regeneration: An Upstream Necessity for Correct Patterning and Brain Formation

[5] Carole Gauron, et al. Sci Rep. Sustained production of ROS triggers compensatory proliferation and is required for regeneration to proceed