| 名称 | H-89 dihydrochloride |
| 描述 | H-89 dihydrochloride (5-Isoquinolinesulfonamide) is a selective inhibitor of cAMP-dependent protein kinase A (PKA) with an IC50 value of 48 nM. It also mildly inhibits PKG, PKC, and casein kinase activity. H-89 dihydrochloride can be used in research areas such as cell proliferation, apoptosis, metabolism, neurotransmission, and endocrine regulation. |
| 细胞实验 | After 48 h in culture, PCl2D cells are cultured in a test medium containing 30 μM H-89 for 1 h and then exposed to a fresh medium that contained both 10 μM forskolin and 30 μM H-89. Cells are scraped off with a rubber policeman and sonicated in the presence of 0.5 mL of 6% trichloroacetic acid. To extract trichloroacetic acid, 2 mL of petroleum ether is added, the preparation mixed and centrifuged at 3000 rpm for 10 min. After aspiration of the upper layer, the residue sample solution is used for determination [1]. |
| 激酶实验 | 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-33P]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 [2]. |
| 动物实验 | H89 (N-[2-(p-Bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide], di-HCl Salt) (10 mg/kg) suspended in 5% DMSO in saline was administered i.p. two hours before each OVA challenge (or two hours before the last OVA challenge). Control animals received equivalent volumes (200 μl) of 5% DMSO in saline [5]. |
| 体外活性 | 方法：HCT116 中加入 1.56-50 μM 浓度梯度的 H-89 dihydrochloride 处理 72 小时，MTT 比色法评估 H-89 dihydrochloride 对细胞活力的影响。
结果：H-89 dihydrochloride 对 HCT116 细胞的生长抑制呈浓度依赖性。[1]
方法：在 LS174T 细胞中，加入 TCF/LEF 荧光素酶报告质粒，转染后加入 20 μM H-89 dihydrochloride 处理 1 小时，结束后，加入 PGE2（浓度 1-10 μM）刺激细胞，继续培养 6 小时。采用 TCF/LEF 荧光素酶报告基因实验来评估 H-89 dihydrochloride 对 Wnt/β-catenin 信号通路转录活性的影响。
结果：在 LS174T 细胞中，20 μM H-89 dihydrochloride 处理能够有效阻断 PGE2 刺激的 TCF/LEF 转录活性。[2] |
| 体内活性 | 方法：成年 SD 大鼠，通过腹腔注射脂多糖(80 μg/kg)建立发热模型。在 LPS 注射前半小时，通过侧脑室埋管注射 H-89 dihydrochloride (0.5， 1.0， 1.5 μg/位点)，以抑制中枢 PKA 活性。注射 LPS 后 4.5 小时 处死取材。
结果：H-89 dihydrochloride 处理显著抑制了 p-TRPV1 的水平，而对总 TRPV1 影响较小且单独给药对正常大鼠的基础体温无显著影响。[3] |
| 存储条件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year
Shipping with blue ice/Shipping at ambient temperature. |
| 溶解度 | H2O : < 1 mg/mL (insoluble or slightly soluble)
DMSO : 104 mg/mL (200.28 mM), Sonication is recommended.
5% DMSO+95% Saline : 3.16 mg/mL (6.09 mM), Solution.
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| 关键字 | S6Kinase | S6K1 | S6 Kinase | Protein kinase inhibitor H-89 | PKA | H-89 Dihydrochloride | H-89 | H89 | H 89 | Autophagy |
| 相关产品 | Guanidine hydrochloride | Naringin | Enzalutamide | Aceglutamide | Alginic acid | Cysteamine hydrochloride | Hemin | Hydroxychloroquine | Sildenafil citrate | Stavudine | Tamoxifen | Paeonol |
| 相关库 | 抑制剂库 | 抗乳腺癌化合物库 | 经典已知活性库 | 已知活性化合物库 | 代谢化合物库 | 激酶抑制剂库 | 抗衰老化合物库 | 血液病分子库 | 免疫/炎症分子化合物库 | 膜蛋白靶向化合物库 | 酪氨酸激酶分子库 | 疼痛相关化合物库 |
