EC-17 (disodium salt) is a folate receptor alpha (FRα) targeting contrast agent with fluorescent properties in the visible light spectrum. The peak excitation and emission wavelengths of EC-17 are 470/520 nm.

Folate receptor alpha^[1]

EC-17 contains the fluorescein fluorochrome and has a spectral wavelength of 490–530 nm. EC-17 is synthesized by a folate (vitamin B9) and fluorescein isothiocyanate (FITC) conjugated through an ethylenediamine spacer to produce folate-FITC, with a molecular weight of 917 kDa. FITC is a derivative of fluorescein functionalized with an isothiocyanate reactive group. The folate-FITC conjugate forms a negatively charged fluorescent molecule that specifically targets cell-surface FRα and is subsequently internalized into the cytoplasm. The signal-to-background ratio (SBR) of EC-17 for HeLa cells range from 0.97 to 7.32 depending on the molarity and concentration of cancer cells^[1].

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.

The mean fluorescence signal from the animals injected with EC-17 to be 42,234 ± 12,234 au^[1]. Fluorescence microscopy for folate-FITC shows a strong signal in all malignant tumors with FR-α expression and no signal in FR-α–negative malignant or benign lesions^[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.

916.82

C₄₂H₃₄N₁₀Na₂O₁₀S

910661-33-5

Room temperature in continental US; may vary elsewhere.

-20°C, sealed storage, away from moisture and light

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

DMSO : 40 mg/mL (43.63 mM; Need ultrasonic)

H₂O : < 0.1 mg/mL (ultrasonic;warming;heat to 60°C) (insoluble)

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 (sealed storage, away from moisture and 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。

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: 1.6 mg/mL (1.75 mM); Suspended solution; Need ultrasonic

  此方案可获得 1.6 mg/mL (1.75 mM) 的均匀悬浊液，悬浊液可用于口服和腹腔注射。

  以 1 mL 工作液为例，取 100 μL 16.0 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: 1.6 mg/mL (1.75 mM); Suspended solution; Need ultrasonic

  此方案可获得 1.6 mg/mL (1.75 mM) 的均匀悬浊液，悬浊液可用于口服和腹腔注射。

  以 1 mL 工作液为例，取 100 μL 16.0 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

• [1]. Tummers QR, et al. Intraoperative imaging of folate receptor alpha positive ovarian and breast cancer using the tumor specific agent EC17. Oncotarget. 2016 May 31;7(22):32144-55.

KB, HeLa, and TC1 cells are plated on a cell culture treated 6-well plate and incubated for 16 hours. Once confluent, EC-17 is added cells. The cells are incubated and sealed in a light-protected environment for 45 minutes. Cells are then washed 3 times with PBS and plated and underwent fluorescence microscopy^[1].

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.

Mice^[1]

Mice are injected subcutaneously in the flank with 1.2×10⁶ TC1 cells (C57BL/6 mice), 1.0×10⁶ HeLa cells (NOD.Cg-Prkdc^scid  Il2rg^tm1Wjl/SzJ mice), or 1.0×106 KB cells (NOD.Cg-Prkdc^scid, Il2rg^tm1Wjl/SzJ mice). Once tumor volume reached approximately 300 mm³ half of the mice are injected with 0.1 mg/kg of EC-17 and the other half with 0.1 mg/kg of OTL38 via tail vein. Three hours later, the fluorescence of tumors is measured using Flocam^[1].

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]. Tummers QR, et al. Intraoperative imaging of folate receptor alpha positive ovarian and breast cancer using the tumor specific agent EC17. Oncotarget. 2016 May 31;7(22):32144-55.