再纯化

The scFv formed inclusion body in E. Coli. 8 mol/L urea was used to solubilize the inclusion body, and the scFv was then purified in two steps , including Ni 2+ chelating affinity chromatography and gel filtration.

大肠杆菌表达的单链抗体包涵体，用8 mol/L尿素裂解，经过Ni离子螯合亲和层析和凝胶过滤两步纯化后，纯度达到97%以上，再通过透析复性除去尿素。

METHODS: Mouse bilateral femur was sterilely isolated. BMSCs were harvested by the Percoll density gradient centrifugation, and purified and proliferated by the adherent method.

无菌分离小鼠双侧股骨，Percoll密度梯度离心法分离骨髓间充质干细胞，再用贴壁筛选法纯化扩增。

Here, by introducing C-terminally 6xHis-tagged human Nap1 gene into Escherichia coli and exploiting the nickel-charged sepharose gel system to purify the human Nap1 protein, we show that the Nap1 enzyme function is preserved in human as in lower organisms such as bacteria and plants using succinamic acid as the substrate.

此研究中，我们将已在胺基末端接上6个组胺酸标记的Nap1基因表现在大肠杆菌中，并使用接合上镍原子的琼脂糖凝胶纯化出人类Nap1蛋白，再以丁醯胺酸作为受质，证明人类Nap1蛋白具有酵素活性。

The serum-free medium is obtained by: the purified human transferring was directly dissolved in the DMEM. Following detoxicating, BSA, cholesterol, insulin and hydrogen peroxidase were directly added in the DMEM, and diluted to a needed concentration.

所用无血清培养基血清替代物是将纯化人转铁蛋白直接溶于DMEM培养基中，再将去毒后的BSA、超声波粉碎后的胆固醇、胰岛素及过氧化氢酶直接加入DMEM培养基中稀释至所需浓度。

For ammonium sulphate precipitation, ammonium sulphate with different concentrations was added to culture filtrate.

再后，论文根据现有实验室的实验条件，选择硫酸铵盐析、凝胶层析和CHT层析对角蛋白酶的粗酶进行了纯化。

METHODS: Umbilical cord blood samples were sterilely isolated using Percoll density gradient centrifugation to harvest intermediate layer cells. DMEM medium containing fetal bovine serum, penicillin, streptomycin and L-glutamine was added. Following several adherences and purification, the floating cells were discarded. Thus, many adherent cells with a confluence were collected. When cells were 60%-80% confluent, cells were digested by trypsin for subculture.

无菌条件下，应用Percoll密度梯度离心法分离脐血标本，收获中间层细胞，加入含胎牛血清、青霉素和链霉素、L-谷氨酰胺的DMEM基础培养液，再经反复贴壁纯化，除去悬浮生长细胞，得到较多呈融合状态的贴壁细胞，待细胞达60%~80%融合时胰酶消化传代。

Methods: The cerebral cortical astrocytes and neurons of rats were cultured and the model of simulated cerebral ischemia and reperfusion was set up in vitro. The activity, survival rate, death rate of the neurons, leakage rate of lactate dehydrogenase and the activity of nitric oxide synthase of cerebral cortical neurons were measured after 18 boors of reperfusion.

分离纯化培养大鼠大脑皮质Ast和神经元，体外模拟脑缺血再灌注损伤模型，用ACM培养损伤后的神经元，测定其神经元的活性、存活率、死亡率、培养液乳酸脱氢酶的漏出率和NOS强阳性细胞的表达量。

By this method, the productivity of 3. 18g/l extracellular polysaccharides was obtained. The optimum extraction method was obtained by RSA. It was determined as follows: 91. 4 ℃, 2. 9h, the weight ratio between mycelia and water was 1 to 3. The productivity was 12. 38% of dry mycelia. Set the purifying methods of Grifola frondosa: Precipitated polysaccharides part by 60% ethanol→removed protein by Sevag method→removed coloring matter by H〓O〓→removed salts by dialyse→DEAE-cellulose column chromatogram→ purified polysaccharides groups.

对于胞内多糖的提取，采用湿菌体经捣碎再高压破壁的方式破碎菌体细胞，采用响应面分析法得出优化后的提取条件为：提取温度91.4℃，提取时间2.9h，料水比1：3，此提取条件下胞内粗多糖的率为干菌体重的12.38％建立了灰树花多糖分离纯化的技术路线 60％乙醇沉淀的粗多糖→Sevag法脱蛋白→H〓O〓脱色→透析脱盐→DEAE-纤维素层析分离→多糖组分。

The eluents were water, 10%, 20 %, 30 % and 50 % ethanol respectively and they flowed through the macroporous resin one by one with five times of the column volume. The obtained eluent of 50 % ethanol was evaporated and the content of total flavonoids was 76 % measured by HPLC.

结果 选择AB-8大孔树脂；最佳纯化工艺为上样液总黄酮质量浓度 60 mg·L-1，pH 5.0，温度 25 ℃；最佳洗脱方式是用5倍柱体积的水及体积分数分别为10 %、20 %、30 %的乙醇依次洗脱，弃去洗脱液后再用5倍柱体积的体积分数为50 %的乙醇洗脱；HPLC法测定总黄酮的含量为76 %。

To examine the hypothesis, we test whether maize beta-amylase can serve as a lipase inhibitor.In order to conduct the interaction assay of beta-amylase and lipase, we adopted a preparative electrophoresis method with a Prep Cell to purify beta-amylase proteins from germinating maize endosperms and soybean cotyledons. We pre-mixed tested proteins with lipid substrate prior to the addition of lipase preparation and assayed the lipid hydrolytic activity.

本实验利用Prep Cell纯化玉米萌芽谷粒和大豆子叶中的beta-淀粉酶，分别将两种不同物种的beta-淀粉酶与胰脂解酶预先反应后，再加入脂解酶反应液中观察活性的变化，结果发现脂解酶的活性未如预期，没有抑制的效果产生，将甘薯beta-淀粉酶以同样方法与胰脂解酶作用，结果同上述两种beta淀粉酶依旧没有抑制的现象。

As she looked at Warrington's manly face, and dark, melancholy eyes, she had settled in her mind that he must have been the victim of an unhappy attachment.

每逢看到沃林顿那刚毅的脸，那乌黑、忧郁的眼睛，她便会相信，他一定作过不幸的爱情的受害者。

Maybe they'll disappear into a pothole.

也许他们将在壶穴里消失