SB202190 (p38 MAPK抑制剂) (RPM0010)

体外活性/In vitro:
1.用SB202190（5-50 μM）处理人Tenon成纤维细胞，MTT法检测细胞活力。SB202190对细胞有毒性，IC50为17.2 μM。[1]
2.用SB202190（5-50 μM）处理人脐静脉内皮细胞HUVEC 6-48 h，Western Blot检测靶点蛋白表达水平。SB202190孵育24小时后，LC3A/B-I转化为PE偶联的LC3A/B-II以浓度依赖的方式增加。[2]

体内活性/In vivo:
1.将SB202190（2 mg/kg）腹腔注射给C57BL/6小鼠，30 min后注射LPS（10 mg/kg），研究p38 MAPK在急性内毒素血症小鼠中的作用。SB202190处理可降低TNF-α水平，显著逆转LPS诱导的左心室抑制，降低诱导的死亡率。[3]
2.将SB202190（5 mg/kg）和OSI-027（10 mg/kg）腹腔注射给携带人CRC肿瘤SW620的BALB/c小鼠，每天一次，持续十天，检测体内抗肿瘤活性。单独使用SB202190增强了SW620异种移植物的肿瘤增殖和肿瘤负荷，联合OSI-027使用显著减弱了异种移植物肿瘤的生长。[4]

激酶实验/Kinase Assay:
All protein kinase activities were linear with respect to time in every incubation. Assays were performed either manually for 10 min at 30°C in 50 μL incubations using [γ-32P]ATP or with a Biomek 2000 Laboratory Automation Workstation in a 96-well format for 40 min at ambient temperature in 25 μL incubations using [γ-32P]ATP. The concentrations of ATP and magnesium acetate were 0.1 mM and 10 mM respectively, unless stated otherwise. This concentration of ATP is 5–10-fold higher than the Km for ATP of most of the protein kinases studied in the present paper, but lower than the normal intracellular concentration, which is in the millimolar range. All assays were initiated with MgATP. Manual assays were terminated by spotting aliquots of each incubation on to phosphocellulose paper, followed by immersion in 50 mM phosphoric acid. Robotic assays were terminated by the addition of 5 μl of 0.5 M phosphoric acid before spotting aliquots on to P30 filter mats. All papers were then washed four times in 50 mM phosphoric acid to remove ATP, once in acetone (manual incubations) or methanol (robotic incubations), and then dried and counted for radioactivity [1].

1.CAS号/CASNO.：152121-30-7|2.化学式/Formula：C20H14FN3O|3.分子量/M.Wt：331.34|4.外观/Appearance：固体|5.溶解度:DMSO: 118.8 mg/mL (358.54 mM), Sonication is recommended.收到重组蛋白产品之后请检查蛋白冻干粉末是否贴于瓶底，如果粉末浮起，开盖之前请先低温离心。将蛋白用说明书中指定的缓冲液复溶至0.1-0.5 mg/mL（请注意蛋白复溶浓度不能低于0.1 mg/mL），室温平衡5-10 min保证充分溶解，复溶过程中请不要剧烈涡旋及吹打蛋白溶液。如需长期储存，建议复溶时添加载体蛋白或者稳定剂（如0.1% BSA, 5% HSA, 10% FBS 或者 5% 海藻糖），同时将复溶后的蛋白溶液按照需求进行分装，储存于-20°C至-80°C，随取随用，避免反复冻融。