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2.8. Fibrinogen Adsorption Test. Fibrinogen adsorption was evaluated using enzyme-linked immunosorbent assay (ELISA) based on the antigen-antibody interaction.36 The fresh whole blood of a healthy volunteer was centrifuged at 3000 rpm to obtain platelet-poor plasma. After being immersed in PBS at 37 C for 3 h, modified and unmodified PHBV films were incubated in plasma at 4 C overnight. Then, all films were blocked with 1% BSA (Sigma-Aldrich) at 37 C for 1 h, followed by reacting with horseradish peroxidase (HRP) conjugated goat antihuman fibrinogen (Sigma-Aldrich) at 37 C for 1 h. Then, TMB (3,30,5,50- tetramethylbenzidine, Sigma-Aldrich) substrate was applied to react with HRP until the solution in the well turned blue. Finally, 2M H2SO4 was added to each well to stop the reaction. The final color of the solution was yellow, which could be read at the 450 nm with a microplate reader. Each type of sample underwent six repeated measurements. 2.9. Statistical Analysis. The results of the cell viability assay and fibrinogen adsorption test were analyzed by ANOVA, and the differences observed between the samples were considered to be statistically significant for P < 0.05. |
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jiangguofeng
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Malyhon: 金币+10, 翻译EPI+1, ★★★★★最佳答案 2015-04-27 14:46:02
Malyhon: 金币+10, 翻译EPI+1, ★★★★★最佳答案 2015-04-27 14:46:02
| 2.8. 纤维蛋白原吸附试验。 根据抗原-抗体交互作用，使用酶联免疫吸附测定(ELISA)对纤维蛋白原吸附进行了评价。36将健康志愿者的新鲜全血以3000rpm（每分钟转数）甩滤得到乏血小板血浆。于37℃的PBS中浸渍３小时之后,修饰和未修饰的PHBV膜于4℃血浆中培育过夜。 然后,所有膜用1% BSA (西格玛-奥德里奇产品）于37℃中封闭1小时,随后37℃与辣根过氧化酶(HRP)结合的羊抗人纤维蛋白原(（西格玛-奥德里奇产品）反应1小时。然后TMB(3,30,5,50 -四甲基联苯胺,西格玛-奥德里奇产品）培养基用于与HRP反应直到容器中的溶液变成蓝色为止。 最后,将2M的H2SO4添加到每个容器中以终止反应。溶液最终的颜色为黄色,该数值可以用酶标仪于450 nm读取。每种样品进行六次反复测量。2.9.统计分析。 细胞活力检测和纤维蛋白原吸附试验通过统计学方差分析进行的，样品之间观测到的差异P<0.05被认为在统计上有显著意义。 |
2楼2015-04-15 10:04:21