Genistein (NPI 031L) is a naturally occurring soy isoflavone, a multi-targeted tyrosine kinase inhibitor. Genistein has antitumor, antioxidant, and anthelmintic properties, and also produces estrogen-like effects in the body.

EGFR:0.6 μM (IC50, Cell Assay)

方法: 乳腺癌细胞 MCF-7 用 Genistein (10-200 µM) 处理 24-48 h，通过 MTT assay 检测细胞活力。
结果: 处理 24 h 后，Genistein 对浓度超过 80 µM 的 MCF-7 细胞具有细胞毒性。用 Genistein 50 µM 以上处理 48 h，在 MCF-7 细胞中诱导细胞毒性。[1]
方法: MCF-7 细胞或分化中的 3T3-L1 细胞用 Genistein (50-200 µM) 处理 24-72 h，通过 Western Blot 检测靶点蛋白表达水平。
结果: MCF-7 细胞中，所有浓度的 Genistein 均下调 ERα 的表达；此外，Genistein 在 48 和 72 h 后的作用更大。与阴性对照 (3T3-L1 前脂肪细胞) 相比，Genistein 处理 48 h 后，3T3-L1 细胞中 ERα 的表达上调了 1.98 倍，以启动分化。[2]

方法: 为检测体内抗肿瘤活性，将 Genistein (2-20 mg/kg in 25 mmol/L Na2CO3) 灌胃给药给携带 B16F10 肿瘤的 B6C3F1 小鼠，每天一次，持续 28 天。
结果: 与 Genistein 的化学预防作用一致，暴露于该化合物显著增加了宿主对 B16F10 肿瘤的抵抗力，这反映在中高剂量水平肿瘤细胞注射后肺肿瘤结节数量的减少上。[3]

Purified FXa is obtained after activation with Russell's viper venom followed by affinity chromatography. The resulting FXa is > 95% pure as judged by sodium dodecylsulfate polyacrylamide gel electrophoresis. The substrate affinity values for FXa, expressed as the Michaelis-Menten-Henri constant (Km), for human, rabbit, rat and dog FXa are determined using the chromogenic substrate S-2765, and are 36, 60, 240 and 70 μM, respectively. The substrate hydrolysis is monitored by measuring absorbance at 405 nm at 25°C for up to 30 min using a SpectraMax 384 Plus plate reader and SoftMax. FXa activity for each substrate and inhibitor concentration pair is determined in duplicate. The Ki values are calculated by non-linear least-squares fitting of the steady-state substrate hydrolysis rates to the equation for competitive inhibition (Equation 1) using GRAFIT, where v equals reactions velocity in OD min?1, Vmax equals maxiumum reaction velocity, S equals substrate concentration, and I equals inhibitor concentration.

Genistein is dissolved in DMSO and stored, and then diluted with appropriate medium before use[1]. The IC50 values for Genistein are determined by the MTT assay. Briefly, the MTT assay is a colorimetric assay that is based on the ability of living but not dead cells to reduce a tetrazolium-based compound to a blue formazan product. The formazan crystals are solubilized in DMSO, and the absorbance is measured at 540 nm. The absorbance at 540 nm is proportional to the number of viable cells. The lC50 values obtained with the MTT assay are compared with the lC50 values obtained by counting viable cells using trypan blue dye exclusion and by tritiated thymidine incorporation into DNA[1].