Isosakuranetin is a flavanone flavonoid which can be found in the fruit of Citrus bergamia.

Isosakuranetin stimulates melanin accumulation in a dose-dependent manner. In addition, Isosakuranetin at 30 μM gradually increases the expression of Tyr, TRP1, and TRP2 after 48 and 72 h. The MTT assay shows that Isosakuranetin decreases the number of cells at concentrations greater than 45 μM. Further analysis of trypan blue staining demonstrates that Isosakuranetin does not affect the viability of B16 melanocytes, but shows that Isosakuranetin decreases their proliferation rate. The tyrosinase activity of B16 melanocytes treated with 15 and 30 μM Isosakuranetin for 72 h is significantly increased by 2- and 3.2-fold, respectively, in comparison with that of the control cells^[1].

Shanghai Jinpan Biotech Co Ltd has not independently confirmed the accuracy of these methods. They are for reference only.

Isosakuranetin dose-dependently increases the paw-withdrawal threshold within the dose range of 1.5 to 6 mg/kg. Post hoc analysis reveals that Isosakuranetin increases the paw-withdrawal threshold and statistical significance is seen at times 30 and 40 min for the dose of 3 mg/kg and 30 to 60 min for the dose of 6 mg/kg^[2].

Shanghai Jinpan Biotech Co Ltd has not independently confirmed the accuracy of these methods. They are for reference only.

286.28

C₁₆H₁₄O₅

480-43-3

异野樱素

Room temperature in continental US; may vary elsewhere.

4°C, sealed storage, away from moisture

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

In Vitro: 

DMSO : 250 mg/mL (873.27 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)。-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    90% corn oil

  Solubility: ≥ 20.83 mg/mL (72.76 mM); Clear solution

  此方案可获得 ≥ 20.83 mg/mL (72.76 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 208.3 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• 2.

  请依序添加每种溶剂： 10% DMSO    40% PEG300    5% Tween-80    45% saline

  Solubility: ≥ 2.08 mg/mL (7.27 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (7.27 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 20.8 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

• 3.

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

  Solubility: ≥ 2.08 mg/mL (7.27 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (7.27 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 20.8 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]. Drira R, et al. Isosakuranetin, a 4′-O-methylated flavonoid, stimulates melanogenesis in B16BL6 murine melanoma cells. Life Sci. 2015 Dec 15;143:43-9.

  [2]. Jia S, et al. Antinociceptive Effects of Isosakuranetin in a Rat Model of Peripheral Neuropathy. Pharmacology. 2017;100(3-4):201-207.

B16 melanoma cells are cultured in 24-well plates at a density of 1×10⁴ cells/well. After 24 h, the cells are incubated for 72 h in the presence of Isosakuranetin at the indicated concentrations. The culture medium is removed and replaced with 500 μL of fresh culture medium containing 10% sterile filtered MTT. After 3 h, the formazan crystals are dissolved in 500 μL/well isopropanol and absorbance is measured at 570 nm against 630 nm. Inhibition of proliferation (%) is expressed as the percentage of viable treated cells in comparison with control cells^[1].

Shanghai Jinpan Biotech Co Ltd has not independently confirmed the accuracy of these methods. They are for reference only.

Male Sprague-Dawley rats weighing 250 to 300 g are habituated to the temperature, humidity, and lighting (12 h light/dark cycle, lights on at 7: 00 a.m.) controlled vivarium and singly housed for at least 1 week before the studies begin. The rotarod test is used to evaluate the potential motor-impairing effect of Isosakuranetin. For the time course studies, baseline measurement is immediately followed by an injection of Isosakuranetin, and then the paw-withdrawal threshold is measured every 15 min until the drug effect dissipates to a point that the paw withdrawal threshold is not significantly different from the predrug data or until 75 min passes^[2].

Shanghai Jinpan Biotech Co Ltd has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Drira R, et al. Isosakuranetin, a 4′-O-methylated flavonoid, stimulates melanogenesis in B16BL6 murine melanoma cells. Life Sci. 2015 Dec 15;143:43-9.

  [2]. Jia S, et al. Antinociceptive Effects of Isosakuranetin in a Rat Model of Peripheral Neuropathy. Pharmacology. 2017;100(3-4):201-207.