Ethoxyquin is an antioxidant which has been used in animal feed for many years and also an inhibitor of heat shock protein 90 (Hsp90).

Ethoxyquin (EQ) does not block Paclitaxel’s (PTX’s) ability to induce cell death in 4 different breast cancer cell lines. Results demonstrate that only when Hsp90 level is down regulated is neuroprotec-tion provided by Ethoxyquin (EQ) no longer seen^[2]. It is found that Ethoxyquin provides neuroprotection with a best efficacy range from 30 to 300 nM. It is interesting to note that higher (μM) concentrations of Ethoxyquin do not provide further neuroprotection and in fact at these higher concentrations Ethoxyquin loses its neuroprotective effects^[3].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

Ethoxyquin (EQ) provides a dose-dependent neuroprotection. Although all 3 doses of Ethoxyquin provide partial neuroprotection against reduction in intraepidermal nerve fiber density, the peak efficacy is with the 75 μg/kg dose^[2]. Rats treated with Ethoxyquin (EQ) alone or control vehicle treated rats gain weight as expected under normal physiological conditions^[3].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

217.31

C₁₄H₁₉NO

91-53-2

乙氧基喹啉

Room temperature in continental US; may vary elsewhere.

Pure form	-20°C	3 years
	4°C	2 years
In solvent	-80°C	6 months
	-20°C	1 month

In Vitro: 

DMSO : ≥ 50 mg/mL (230.09 mM)

H₂O : < 0.1 mg/mL (insoluble)

* “≥” means soluble, but saturation unknown.

*

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。-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: ≥ 3.5 mg/mL (16.11 mM); Clear solution

  此方案可获得 ≥ 3.5 mg/mL (16.11 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 35.0 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加入50 μL Tween-80，混合均匀；然后继续加入 450 μL生理盐水定容至 1 mL。

  将 0.9 g 氯化钠，完全溶解于 100 mL ddH₂O 中，得到澄清透明的生理盐水溶液

• 2.

  请依序添加每种溶剂： 10% DMSO    90% (20% SBE-β-CD in saline)

  Solubility: ≥ 3.5 mg/mL (16.11 mM); Clear solution

  此方案可获得 ≥ 3.5 mg/mL (16.11 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 35.0 mg/mL 的澄清 DMSO 储备液加到 900 μL 20% 的 SBE-β-CD 生理盐水水溶液中，混合均匀。

  将 2 g 磺丁基醚 β-环糊精加入 5 mL 生理盐水中，再用生理盐水定容至 10 mL，完全溶解，澄清透明
• 3.

  请依序添加每种溶剂： 10% DMSO    90% corn oil

  Solubility: ≥ 3.5 mg/mL (16.11 mM); Clear solution

  此方案可获得 ≥ 3.5 mg/mL (16.11 mM，饱和度未知) 的澄清溶液，此方案不适用于实验周期在半个月以上的实验。

  以 1 mL 工作液为例，取 100 μL 35.0 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• [1]. Sadikot T, et al. Development of a high-throughput screening cancer cell-based luciferase refolding assay for identifying Hsp90 inhibitors. Assay Drug Dev Technol. 2013 Oct;11(8):478-88.

  [2]. Zhu J, et al. Ethoxyquin prevents chemotherapy-induced neurotoxicity via Hsp90 modulation. Ann Neurol. 2013 Dec;74(6):893-904.

  [3]. Zhu J, et al. Ethoxyquin provides neuroprotection against cisplatin-induced neurotoxicity. Sci Rep. 2016 Jun 28;6:28861.

To evaluate the impact of Ethoxyquin (EQ) on Paclitaxel’s (PTX’s) ability to kill breast cancer cells, conditions for culturing 4 cancer cell lines and measuring the ATP levels are optimized for the 96-well plate format. Briefly, 1,500 cells/well in media are plated in 96-well plates for 24 hours. Constant concentrations of PTX with or without Ethoxyquin are added to the wells for another 24 hours. Cellular ATP levels are measured using kit according tomanufacturer ’s protocol^[2].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

Briefly, DRGs are harvested from embryonic day 14.5 rats according to standard protocols, then cells are plated onto collagen-coated glass coverslips and allowed to extend neurites for 24 hours in media (Neurobasal medium, 50 mM pencillin-streptomycin (PS), 0.2% FBS, 0.5 mM glutamine, 1×B-27 supplement, 0.2% glucose,10 ng/mL glial cell line-derived neurotrophic factor). Ethoxyquin (EQ) or vehicle control is added to the wells for another 24-hour incubation. DRG cells are fixed with 4% paraformaldehyde and stained with anti-βIII-tubulin antibody to delineate the axons. Axon lengths are measured in multiple fields using a random sampling method^[2].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

To examine the effect of Ethoxyquin (EQ) on Paclitaxel’s (PTX’s) ability to reduce tumor burden in vivo, a mouse model in which breast cancer cell line SUM-159 (3×10⁶ tumor cells suspended in phosphate-buffered saline in a final volume of 0.15 mL) is injected subcutaneously into adult male nude mice. When the tumor size reaches 5 mm in diameter, the animals are randomly assigned to PTX or PTX with Ethoxyquin groups. Ethoxyquin is given by intraperitoneal administration on a daily basis for 3 weeks. At the end of 3 weeks, animals are euthanized and tumor size and weight are measured (n=5 per group)^[2].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Sadikot T, et al. Development of a high-throughput screening cancer cell-based luciferase refolding assay for identifying Hsp90 inhibitors. Assay Drug Dev Technol. 2013 Oct;11(8):478-88.

  [2]. Zhu J, et al. Ethoxyquin prevents chemotherapy-induced neurotoxicity via Hsp90 modulation. Ann Neurol. 2013 Dec;74(6):893-904.

  [3]. Zhu J, et al. Ethoxyquin provides neuroprotection against cisplatin-induced neurotoxicity. Sci Rep. 2016 Jun 28;6:28861.