Hydroethidine也称为Dihydroethidium，是一种最常用的超氧化物阴离子荧光检测探针,能有效地检测活性氧类。这种染料可以自由地进入细胞，在细胞内超氧化物阴离子作用下脱氢形成溴化乙锭。溴化乙锭可以与RNA或DNA结合产生红色荧光。当细胞内的超氧化物阴离子水平较高时，产生的溴化乙锭较多，红色荧光就较强，反之则较弱。这样就可以用二氢乙锭进行超氧化物阴离子水平的检测。
该探针已经广泛地用于NK细胞、中性粒细胞、内皮细胞、细胞HL60和巨噬细胞的研究，并且作为一种重要的染料用于鉴定肿瘤中细胞的增殖和低氧性。
| 产品名称（Product Name） | Dihydroethidium (Hydroethidium) |
| CAS号（CAS NO.） | 38483-26-0 |
| 分子式（Molecular Formula） | C21H21N3 |
| 分子量（Molecular Weight） | 315.41 |
| 激发和发射波长（Ex/Em） | 518 nm/610 nm |
| 纯度( Purity ) | ≥90% |
| 外观（Appearance） | 红色粉末 |
| 溶解性（Solubility） | 溶于DMSO |
| 结构式（Structure） |
注意事项
1）请勿吸入、吞咽或者直接接触皮肤和眼睛。
2）二氢乙锭容易氧化且见光易分解，一定要避光干燥保存。
3）在没有细胞和外界诱导剂存在的情况下，溶解在培养液或者缓冲溶液中（PBS/ HBSS/ HEPES）的染料会发生水解，被空气或通过光诱导的方式氧化，而使溶液的荧光强度增加。
4）为了您的安全和健康，请穿实验服并戴一次性手套操作。
5）本产品仅用于科研用途，禁止用于人身上。
使用方法
1. 工作液的配制
本品以粉末形式提供，使用前需要经过短暂离心，用高纯度的DMSO制备成5~10 mM储液。分装后避光冻存于-20℃。使用前用缓冲液（PBS/ HBSS/ HEPES）稀释母液至所需工作浓度，通常5~20 μM就可以满足实验需求，具体的浓度要根据实验需要进行进一步的调整。
2. 染色
将细胞和染色液按照1:1的比例进行混合（例如，100 μL的细胞悬液加入100 μL的染色工作液），室温或37℃孵育60 min。
3. 荧光显微镜观察
二氢乙锭本身为蓝色荧光，最大激发波长为370 nm，最大发射波长为420 nm，脱氢后与RNA或DNA结合产生红色荧光，最大激发波长为300 nm，最大发射波长为610 nm，实际观察时也可以使用518 nm作为激发波长。
客户使用本产品发表的科研文献（部分）
[1] Feng L, et al. Neutrophil-like Cell-Membrane-Coated Nanozyme Therapy for Ischemic Brain Damage and Long-Term Neurological Functional Recovery. ACS Nano. 2021 Feb 23;15(2):2263-2280. doi: 10.1021/acsnano.0c07973. Epub 2021 Jan 11. PMID: 33426885. IF: 15.881
[2] [1] Rlabc D, et al. Inhibition of ROS activity by controlled release of proanthocyanidins from mesoporous silica nanocomposites effectively ameliorates heterotopic tendon ossification[J]. Chemical Engineering Journal, 2021. IF: 13.273
[3] Feng J, et al. AKAP1 contributes to impaired mtDNA replication and mitochondrial dysfunction in podocytes of diabetic kidney disease. Int J Biol Sci. 2022 Jun 13;18(10):4026-4042. doi: 10.7150/ijbs.73493. PMID: 35844803. IF: 10.75
[4] Chen L, et al. Therapeutic effect of SIRT3 on glucocorticoid-induced osteonecrosis of the femoral head via intracellular oxidative suppression. Free Radic Biol Med. 2021 Nov 20; 176:228-240. PMID: 34260898. IF: 7.376
[5] Qiao S, et al. Bergenin impedes the generation of extracellular matrix in glomerular mesangial cells and ameliorates diabetic nephropathy in mice by inhibiting oxidative stress via the mTOR/β-TrcP/Nrf2 pathway[J]. Free Radical Biology and Medicine, 2019, 145: 118-135. IF: 7.376
[6] Che Y, et al. The Combination of Rhodosin and MMF Prolongs Cardiac Allograft Survival by Inhibiting DC Maturation by Promoting Mitochondrial Fusion. Oxid Med Cell Longev. 2022 Jul 9; 2022:7260305. PMID: 35855862. IF: 7.31
[7] Hu J, et al. Inhibition of CACNA1H attenuates doxorubicin-induced acute cardiotoxicity by affecting endoplasmic reticulum stress[J]. Biomedicine & Pharmacotherapy, 2019, 120: 109475. IF: 6.529
[8] Lin Z, et al. Eicosapentaenoic Acid-Induced Autophagy Attenuates Intervertebral Disc Degeneration by Suppressing Endoplasmic Reticulum Stress, Extracellular Matrix Degradation, and Apoptosis. Front Cell Dev Biol. 2021 Nov 4; 9:745621. doi: 10.3389/fcell.2021.745621. PMID: 34805156; PMCID: PMC8599281. IF: 6.684
[9] Chen L, et al. Apelin-13 induces mitophagy in bone marrow mesenchymal stem cells to suppress intracellular oxidative stress and ameliorate osteoporosis by activation of AMPK signaling pathway. Free Radic Biol Med. 2021 Feb 1; 163:356-368. doi: 10.1016/j.freeradbiomed.2020.12.235. Epub 2020 Dec 30. PMID: 33385540. IF: 6.17
[10] Li D, et al. Cereulide Exposure Caused Cytopathogenic Damages of Liver and Kidney in Mice. Int J Mol Sci. 2021 Aug 24;22(17):9148. doi: 10.3390/ijms22179148. PMID: 34502057; PMCID: PMC8431326. IF：5.924
[11] Pan B, et al. Azilsartan Suppresses Osteoclastogenesis and Ameliorates Ovariectomy-Induced Osteoporosis by Inhibiting Reactive Oxygen Species Production and Activating Nrf2 Signaling. Front Pharmacol. 2021 Nov 26;12:774709. doi: 10.3389/fphar.2021.774709. PMID: 34899338; PMCID: PMC8662525. IF：5.811
[12] Wang K, et al. Icariin Prevents Extracellular Matrix Accumulation and Ameliorates Experimental Diabetic Kidney Disease by Inhibiting Oxidative Stress via GPER Mediated p62-Dependent Keap1 Degradation and Nrf2 Activation. Front Cell Dev Biol. 2020 Jul 17; 8:559. doi: 10.3389/fcell.2020.00559. PMID: 32766240; PMCID: PMC7379398. IF：5.186
[13] Zhu L, et al. The critical role of RasGRP4 in the growth of diffuse large B cell lymphoma. Cell Commun Signal. 2019 Aug 13;17(1):92. doi: 10.1186/s12964-019-0415-6. PMID: 31409422; PMCID: PMC6693169. IF：5.128
冰袋运输。粉末直接保存于-20℃，有效期2年。产品易被氧化，建议分装后-20℃避光密封保存，避免反复冻融，避光，保持干燥。
Q：探针可以直接用含血清的培养液稀释吗？
A：可以。
Q：使用流式细胞仪探测的话选择什么通道？
A：PE或FL2通道。
Q：用荧光显微镜检测荧光，需要抗淬灭剂封片吗？
A：加少量PBS即可观察，不需要抗淬灭剂封片。
Q：多聚甲醛固定后包埋的冰冻组织样本切片可以用吗？
A：不合适，这款探针用于检测活细胞。
Q：我们的探针是否适用于检测溶液中的超氧阴离子？
A: 不适用，我们的探针只适合做细胞实验。
Q: 二氢乙锭(超氧化物阴离子荧光探针)，这个探针可以做冰冻切片吗?
A: 这个探针是染活细胞的，未做过冰冻切片的验证，不推荐。
[1] Feng L, Dou C, Xia Y, et al. Neutrophil-like Cell-Membrane-Coated Nanozyme Therapy for Ischemic Brain Damage and Long-Term Neurological Functional Recovery. ACS Nano. 2021;15(2):2263-2280. doi:10.1021/acsnano.0c07973(IF:15.881)
[2] Feng J, Chen Z, Ma Y, et al. AKAP1 contributes to impaired mtDNA replication and mitochondrial dysfunction in podocytes of diabetic kidney disease. Int J Biol Sci. 2022;18(10):4026-4042. Published 2022 Jun 13. doi:10.7150/ijbs.73493(IF:10.750)
[3] Chen L, Wang BZ, Xie J, et al. Therapeutic effect of SIRT3 on glucocorticoid-induced osteonecrosis of the femoral head via intracellular oxidative suppression. Free Radic Biol Med. 2021;176:228-240. doi:10.1016/j.freeradbiomed.2021.07.016(IF:7.376)
[4] Lin Z, Ni L, Teng C, et al. Eicosapentaenoic Acid-Induced Autophagy Attenuates Intervertebral Disc Degeneration by Suppressing Endoplasmic Reticulum Stress, Extracellular Matrix Degradation, and Apoptosis. Front Cell Dev Biol. 2021;9:745621. Published 2021 Nov 4. doi:10.3389/fcell.2021.745621(IF:6.684)
[5] Chen L, Shi X, Xie J, et al. Apelin-13 induces mitophagy in bone marrow mesenchymal stem cells to suppress intracellular oxidative stress and ameliorate osteoporosis by activation of AMPK signaling pathway. Free Radic Biol Med. 2021;163:356-368. doi:10.1016/j.freeradbiomed.2020.12.235(IF:6.170)
[6] Li D, Lin R, Xu Y, et al. Cereulide Exposure Caused Cytopathogenic Damages of Liver and Kidney in Mice. Int J Mol Sci. 2021;22(17):9148. Published 2021 Aug 24. doi:10.3390/ijms22179148(IF:5.924)
[7] Pan B, Zheng L, Fang J, et al. Azilsartan Suppresses Osteoclastogenesis and Ameliorates Ovariectomy-Induced Osteoporosis by Inhibiting Reactive Oxygen Species Production and Activating Nrf2 Signaling. Front Pharmacol. 2021;12:774709. Published 2021 Nov 26. doi:10.3389/fphar.2021.774709(IF:5.811)
[8] Wang K, Zheng X, Pan Z, et al. Icariin Prevents Extracellular Matrix Accumulation and Ameliorates Experimental Diabetic Kidney Disease by Inhibiting Oxidative Stress via GPER Mediated p62-Dependent Keap1 Degradation and Nrf2 Activation. Front Cell Dev Biol. 2020;8:559. Published 2020 Jul 17. doi:10.3389/fcell.2020.00559(IF:5.186)
[9] Zhu L, Xia C, Wu L, et al. The critical role of RasGRP4 in the growth of diffuse large B cell lymphoma. Cell Commun Signal. 2019;17(1):92. Published 2019 Aug 13. doi:10.1186/s12964-019-0415-6(IF:5.111)
[10] Wu Q, Hong J, Wang Z, et al. Abnormal Ribosome Biogenesis Partly Induced p53-Dependent Aortic Medial Smooth Muscle Cell Apoptosis and Oxidative Stress. Oxid Med Cell Longev. 2019;2019:7064319. Published 2019 May 9. doi:10.1155/2019/7064319(IF:4.868)
[11] Hu J, Wu Q, Wang Z, et al. Inhibition of CACNA1H attenuates doxorubicin-induced acute cardiotoxicity by affecting endoplasmic reticulum stress. Biomed Pharmacother. 2019;120:109475. doi:10.1016/j.biopha.2019.109475(IF:3.743)
[12] Li X, Ding D, Yao J, et al. Chromatin remodeling factor BAZ1A regulates cellular senescence in both cancer and normal cells. Life Sci. 2019;229:225-232. doi:10.1016/j.lfs.2019.05.023(IF:3.448)
[13] Zhang W, Dun Y, You B, et al. Trimetazidine and exercise offer analogous improvements to the skeletal muscle insulin resistance of mice through Nrf2 signaling. BMJ Open Diabetes Res Care. 2022;10(2):e002699. doi:10.1136/bmjdrc-2021-002699(IF:3.388)
[14] Ma H, Song X, Huang P, et al. Myricetin protects natural killer cells from arsenite induced DNA damage by attenuating oxidative stress and retaining poly(ADP-Ribose) polymerase 1 activity. Mutat Res Genet Toxicol Environ Mutagen. 2021;865:503337. doi:10.1016/j.mrgentox.2021.503337(IF:2.873)
