Question

求大神帮忙翻译一下，急用

2.5. Determination of protein profile and molecular weight
Denaturing electrophoresis (SDS–PAGE) was performed according to Bizani et al. (2005). A discontinuous system was used consisting of two acrylamide gels, a separating gel (15% acrylamide) and a stacking gel (4% acrylamide), and a running buffer (Tris– Glycine–SDS). For the preparation of the samples 5 lg of each extract were taken and diluted in 6 ll of loading buffer 5(5) distilled water was added to achieve a final volume of 50 m. The samples were incubated for 5 min at 95 C in order to favour the denaturation of proteins. Gels were run at 100 V for 2 h at room temperature. The gels were incubated for 2 h in a solution of 25% methanol, 10% acetic acid and 0.03% formaldehyde to fix the proteins. Then the gels were washed 3 times for 20 min with 30% ethanol. The washed gels were incubated in dark for 1 min in a solution of 0.01% sodium thiosulfate. To remove the remaining sodium thiosulfate, the gel is rinsed three times in distilled water and incubated for 20 min in darkness in a solution of 0.2% of silver nitrate and 0.14% formaldehyde; the silver solution was removed with three washings of distilled water. The development was conducted with 6.0% sodium carbonate, 0.03% formaldehyde and 0.03% sodium thiosulfate; the gel was stirred until banding watch it, the reaction is stopped with a solution of 25% methanol and 10% acetic acid. The relative molecular weight of the polypeptides of honey samples was determined by plotting a standard curve of Rf vs. logMw for known samples (ladder), and read off the logMw of the sample after measuring distance migrated on the same gel.

Answer 1

2.5。蛋白质含量和分子量的测定
变性凝胶电泳（SDS–页）是根据bizani等人进行的。（2005）。不连续系统由两丙烯酰胺凝胶，分离凝胶（15%丙烯酰胺）和浓缩胶（4%丙烯酰胺），和一个运行缓冲液（三–甘氨酸–SDS）。对每个提取的样品制备5 LG被稀释在加载缓冲区5会6（5）加入蒸馏水，达到最后一卷50米的样品孵育5分钟，在95℃以利于蛋白质的变性。凝胶运行在100 V，在室温下的2小时。的凝胶孵育2小时，在溶液中的25%的甲醇，10%乙酸和0.03%的甲醛来固定的蛋白质。然后用30%乙醇洗涤3次，用20分钟。洗涤凝胶孵育1分钟，在一个黑暗的0.01%硫代硫酸钠溶液。去除剩余的硫代硫酸钠，凝胶冲洗三次蒸馏水和孵育20分钟在黑暗中硝酸银和0.14% 0.2%甲醛溶液；银溶液用三次蒸馏水去除。发展是用6%碳酸钠溶液进行的，0.03%的甲醛和0.03%硫代硫酸钠；凝胶搅拌直到带看，反应和25%甲醇和10%的乙酸溶液停止。通过一个标准的Rf对已知样本logmw曲线绘制测定蜂蜜样品的多肽的相对分子量（阶梯），然后读取样品的logmw后测量距离在同一凝胶迁移。
望采纳，谢谢。