Busulfan is a potent alkylator with selective immunosuppressive effect on bone marrow.

DNA Alkylator^[1]

Busulfan is a potent alkylating agent, and used as a chemotherapeutic agent. Busulfan (120 μM) results in premature senescence in WI38 cells via the Erk and p38 MAPK pathway, reduces GSH and increases ROS production, but the production can be suppressed by NADPH oxidase^[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.

Busulfan (40 mg/kg, i.p) increases apoptosis and decreases the testis weight in mice. The testes of Busulfan-treated mice exhibits higher level of pRB expression, inhibits Rb phosphorylation and PCNA expression compared to the control^[2]. Busulfan (2.5, 5.0 mg/kg, i.p.) causes earlier occurrence of persistent esturs in a dose dependent manner in rats. Busulfan (5.0 mg/kg) also increases the incidence of uterine adenocarcinomas and multiplicity of uterine neoplastic lesions. In addition, Busulfan decreases the serum 17β-estradiol(E2), progesterone and inhibin levels in rats, and elevates the E2/progesterone ratio only at 5.0 mg/kg^[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.

246.30

C₆H₁₄O₆S₂

55-98-1

白消安

Room temperature in continental US; may vary elsewhere.

DMSO : 50 mg/mL (203.00 mM; Need ultrasonic)

H₂O : 1 mg/mL (4.06 mM; ultrasonic and warming and heat to 80°C)

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。

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: ≥ 2.5 mg/mL (10.15 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (10.15 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 25.0 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 (8.44 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (8.44 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 (8.44 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (8.44 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]. Probin V, et al. Busulfan-induced senescence is dependent on ROS production upstream of the MAPK pathway. Free Radic Biol Med. 2007 Jun 15;42(12):1858-65. Epub 2007 Mar 31.

  [2]. Choi YJ, et al. Murine male germ cell apoptosis induced by busulfan treatment correlates with loss of c-kit-expression in a Fas/FasL- and p53-independent manner. FEBS Lett. 2004 Sep 24;575(1-3):41-51.

  [3]. Yoshida M, et al. Reduction of primordial follicles caused by maternal treatment with busulfan promotes endometrial adenocarcinoma development in donryu rats. J Reprod Dev. 2005 Dec;51(6):707-14. Epub 2005 Sep 22.

WI38 cells are seeded in wells of a 24-well plate at 60,000 cells/well and cultured overnight. They are treated with Busulfan for 24 h, after which the drug is removed and the cells are cultured in fresh medium for up to 11 d. Untreated cells are used as control. Every 3 d the cells are trypsinized, counted and sub-cultured in a fresh medium at 1:5 and 1:2 dilutions for control untreated and Busulfan-treated cells, respectively^[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^[2]
One hundred twenty ICR male mice ranging in age from 8 to 12 weeks (30-40 g) are used. Eight-week-old ICR male mice receive a single intraperitoneal injection of Busulfan (40 mg/kg) diluted in sesame oil. The mice are housed in wire cages at 22 ± 1°C under a 12-h light-dark cycle with 70% humidity and fed ad libitum^[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]. Probin V, et al. Busulfan-induced senescence is dependent on ROS production upstream of the MAPK pathway. Free Radic Biol Med. 2007 Jun 15;42(12):1858-65. Epub 2007 Mar 31.

  [2]. Choi YJ, et al. Murine male germ cell apoptosis induced by busulfan treatment correlates with loss of c-kit-expression in a Fas/FasL- and p53-independent manner. FEBS Lett. 2004 Sep 24;575(1-3):41-51.

  [3]. Yoshida M, et al. Reduction of primordial follicles caused by maternal treatment with busulfan promotes endometrial adenocarcinoma development in donryu rats. J Reprod Dev. 2005 Dec;51(6):707-14. Epub 2005 Sep 22.