Recilisib (ON 01210) is a radioprotectant, which can activate AKT, PI3K activities in cells^[1].

Recilisib Sodium (up to 50 µM) shows a normal distribution of cells throughout the cell cycle, with a slight reduction in the number of cells in S-phase at 50 µM. Continuous exposure of Recilisib Sodium (100 µM) does not result in cell death. Recilisib Sodium does not inhibit the ability of human bone marrow to form colonies in methylcellulose at either timepoint. Recilisib Sodium treatment does not inhibit the colony forming potential of human bone marrow cells. Recilisib Sodium provides dose dependent protection of human bone marrow cells at all three doses of IR. Recilisib Sodium activates the phosphorylation of AKT and GSK3α/β in HFL cells. Recilisib Sodium increases PI3K activity in HFL-1 cells and murine bone marrow cells in response to radiation exposure. Recilisib Sodium treatment in combination with radiation alters the MAPK signaling pathway^[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.

Recilisib Sodium (500 mg/kg) significantly increases the rate of recovery and differentiation of primitive bone marrow myeloid progenitor cells in mice. Recilisib Sodium in combination with radiation reduces CFU numbers in mice, but the Recilisib Sodium-treated mice consistently retain a capability to form differentiated colonies. Recilisib Sodium treated mice have a progenitor cell population that is never completely depleted by radiation exposure^[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.

336.79

C₁₆H₁₃ClO₄S

334969-03-8

Room temperature in continental US; may vary elsewhere.

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

In Vitro: 

DMSO : 35.71 mg/mL (106.03 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。-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: ≥ 2.08 mg/mL (6.18 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (6.18 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 20.8 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• [1]. Kang AD, et al. ON01210.Na (Ex-RAD) mitigates radiation damage through activation of the AKT pathway. PLoS One. 2013;8(3):e58355.

PI3-kinase assays are performed using exponentially growing HFL-1 or freshly harvested murine bone marrow cells that are treated with increasing concentrations of Recilisib Sodium for 2 hours and then irradiated with 10 Gy IR. These cells are then returned to the incubator for 2 to 24 hours and lysed in HEPES pH 7.5 lysis buffer. PI-3K is immunoprecipitated using an anti-PI3 Kinas polyclonal antibody for 2 hours at 4°C. Protein A/G PLUS-Agarose is incubated with immunoprecipitates for 8-16 hours at 4°C and the resulting immunoprecipitates washed with twice HEPES pH 7.5 lysis buffer and once with the kinase buffer (20 mM Tris pH 7.5, 1mM EGTA, 10mM MgCl₂, 2 mM DTT, 0.01% NP-40). L-α-Phosphatidylinositol (12.5 mM) and ATP (10 µM) are added to the kinase buffer (60 µL per sample) and incubated at 30°C for 30 minutes. The reaction is stopped by addition of 100 µL of 1N HCl and extracted by addition of 200 µL CHCl₃/CH₃OH (1:1). The extracted samples are vortexed, centrifuged and the lower organic phases containing phospholipids are dried at 27°C for 2 hours. The dried samples are resuspended in 10 µL of PI-4-P standard (0.5 mL CHCl₃, 0.5 mL CH₃OH, 2.5 µL HCl) and spotted on TLC plates (VWR). The spotted plate is subjected to thin layer chromatography in CHCl₃/CH₃OH/NH4OH (40:40:15). The TLC plate is dried and subjected to autoradiography.

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.

For cytotoxicity assays, cells (1.0×10⁵ cells/mL HPGM) are treated with various concentrations of Recilisib Sodium or vehicle without radiation treatment for 2 or 24 hours. The cells are washed and plated into methocult using gridded dishes. The total number of colony forming units (CFUs) is determined 14 days post-plating by microscopic observation using an Olympus IMT-2 microscope.

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]. Kang AD, et al. ON01210.Na (Ex-RAD) mitigates radiation damage through activation of the AKT pathway. PLoS One. 2013;8(3):e58355.