Sanguinarine (Sanguinarin) chloride, a benzophenanthridine alkaloid derived from the root of Sanguinaria Canadensis, can stimulate apoptosis via activating the production of reactive oxygen species (ROS). Sanguinarine-induced apoptosis is associated with the activation of JNK and NF-κB.

Apoptosis^[1]

Sanguinarine (SANG)-induced apoptosis is associated with the activation of JNK and NF-κB signal pathways.To determine the effects of Sanguinarine on cell viability, 22B-cFluc cells are stimulated with different concentrations of Sanguinarine for 24 h, and then a CKK-8 assay is performed. The treatment with Sanguinarine decreases the proliferation of 22B cells in a dose-dependent manner. Meanwhile, the cytosolic extracts of 22B-cFluc cells treated with different dose of Sanguinarine are measured to detect cellular caspase-3 activity using Ac-DEVD-pNA, which is a validated caspase-3 substrate. The absorbance at 450 nm increases in a dose-dependent manner, indicating increased caspase-3 activity stimulated by Sanguinarine^[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.

To evaluate the apoptosis induced by Sanguinarine (SANG) in vivo, 22B-cFluc cells are inoculated subcutaneously into one flank of nude mice and xenograft models are allowed to establish. Mice are treated intravenously with 10 mg/kg of Sanguinarine. At 24, 48 and 72 h after treatment, bioluminescent imaging is performed after i.p. injection of mice with 150 mg/kg of D-luciferin substrate. Sanguinarine treatment induces an obvious increase of luminescent signal as early as 48 h after initial treatment. A sustained bioluminescent imaging (BLI) intensity increased is observed throughout the course of experiment. At 72 h after treatment, the tumors are collected and subjected to TUNEL staining for evaluating apoptosis. Compared with the control tumors, the group treated with Sanguinarine exhibits significantly more cell apoptosis, indicated by the increased green signals from the sporadic apoptotic 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.

367.78

C₂₀H₁₄ClNO₄

5578-73-4

氯化血根碱

Room temperature in continental US; may vary elsewhere.

4°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)

In Vitro: 

H₂O : 6 mg/mL (16.31 mM; Need ultrasonic)

DMSO : 5 mg/mL (13.60 mM; Need ultrasonic)

*

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。

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: ≥ 0.5 mg/mL (1.36 mM); Clear solution

  此方案可获得 ≥ 0.5 mg/mL (1.36 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 5.0 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: ≥ 0.5 mg/mL (1.36 mM); Clear solution

  此方案可获得 ≥ 0.5 mg/mL (1.36 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 5.0 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]. Wang Y, Noninvasive bioluminescence imaging of the dynamics of sanguinarine induced apoptosis via activation of reactive oxygen species. Oncotarget. 2016 Apr 19;7(16):22355-67.

The caspase-3 activity is measured using a caspase-3 activity assay kit. Briefly, the cells treated by different concentrations of Sanguinarine (0.5 μM, 1 μM, 2 μM, 4 μM) or control DMSO are collected, washed and lysed in a lysis buffer for 30 min on ice. The supernatants are then collected by centrifuging at 1,2000 rpm for 10 min. The Ac-DEVD-pNA (2 mM) is added to each sample and incubated at 37°C for 1 h. The optical density (OD) of each sample is finally quantified at a wavelength of 405 nm using a spectrophotometer. The p-NA standard is used to calibrate the caspase-3 activity of each sample^[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 cell viability of Sanguinarine is determined by CCK-8 assay using a cell counting kit-8. Briefly, 22B-cFluc cells are seeded in a 96-well plate (5×10³ cells/well) and treated with different concentrations of Sanguinarine (0.5 μM, 1 μM, 2 μM, 4 μM) for 24 h. Then 10 mL CKK-8 is added to each well for 4 h and the absorbance at 450 nm is measured with a microplate reader. The optical density (OD) values are determined to reflect the viable cell populations from each well^[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]
Xenografted tumor models are prepared by injection of 1×10⁷ 22B-cFluc cells suspended in PBS into nude mouse (n=6). After tumors reach a volume of approximately 100 mm³, Sanguinarine (10 mg/kg) is i.v. injected into mice. After injection for 24 h, 48 h and 72 h, mice are given a single i.p. dose of 150 mg/kg D-luciferin and bioluminescence imaging are performed using a Xenogen Lumina II system. The signal intensity in the region of interest is expressed using the Living Image software 4.1. For the anti-tumor therapy studies, one group of tumor-bearing mice (n=6) receive intravenously 10 mg/kg of Sanguinarine every other day throughout the experimental period, while the control group of mice (n=6) receive DMSO only. Tumor growth measurement is calculated^[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]. Wang Y, Noninvasive bioluminescence imaging of the dynamics of sanguinarine induced apoptosis via activation of reactive oxygen species. Oncotarget. 2016 Apr 19;7(16):22355-67.