蛋白质组学

Sepharose的说明书操作部分的翻译,请指教!

1.  Equilibrate all material to the temperature at which the chromatography will be performed.将所有材料平衡至进行层析的温度。
2.  Prepare the gel as described above and de-gas the gel slurry.将凝胶如上预备,并脱气。
3.  Mount a packing reservior at the top of a column and rinse with distilled water.将预装的储液容器置于柱的顶部,以蒸馏水清洗。
4.  Insert an end piece or an adaptor at the bottom of the column.将柱底部的末端或接头插入。
5.  Mount the column with the packing reservoir vertically on a laboratory stand. Eliminate air from the column dead sapces by flushing the end piece or adaptor with eluent or distilled water. Make sure no air has been trapped under the net. Close the column outler with approximately one centimeter of eluent remaining in the column.将柱与储液器垂直安装在试管架上。以洗脱液或蒸馏水冲洗将空气从柱的死体积中排出,确保没有空气余留在网下。 将柱的出口关闭,在柱中保留约一厘米的洗脱液。
6.  Pour the slurry in a single operation. Pouring the slurry down a glass rod held against the wall of the column will minimize the introduction of air bubbles. Fill the reservoir to the top with eluent or distilled water.将凝胶浆体一次性倒入。沿抵着柱壁的玻棒倒入凝胶浆可以减少混入气泡。在储液器装入洗脱液或蒸馏水。
7.  Screw on the reservoir cap tightly and connect it to the pump.拧紧贮液器盖,与泵连接。
8.  Open the bottom outlet of the column and start the pump at the flow given for step 1in Table 2. 打开底部的出口,将泵以表2中的步骤1给出的流率启动。
9.  Pack the gel until the gel bed has reached a constant height.填压凝胶直至凝胶床体到达一个恒定的高度。
10.  Stop the pump, close the column outlet and remove the packing reservoir. This is most easily done by first removing the column from the stand and then unscrewing the reservoir over a sink.停止泵压,关闭柱的出口,去除填压用的贮液器。首先从试管架上卸下柱体,然后从槽上拧下贮液器,这样就很容易完成。
11.  Re-mount the column on the stand. Carefully fill the rest of the column with eluent to form an upward menisus. 在试管架上重新装上层析柱。以洗脱液仔细地填满柱的余留部分,形成一个向上的半月形。
12.  Insert the adaptor at an angle into the column, ensuring that no air is trapped under the net.以一定角度插入接头,保证网下没有空气。
13.  Slide the plunger slowly down the column so that the air above the net and in the capillary tubings is displaced by eluent. Valves on the inlet side of the column should be turned in all directions during this procedure to ensure that air is removed.将接头的活塞缓缓滑入层析柱,这样在网上和在毛细管中的空气的可以被洗脱液置换。这个过程入口侧的阀应在各个方向内旋,保证去除空气。
14.  Lock the adaptor in position on the gel surface. Open the column outlet and apply a constant pressure (flow for Sepharose 2B) given for step 2 in Table 2, until the gel bed has reached a constant height.将接头固定在凝胶表面位置,打开柱的出口,以表2中的步骤2恒压(适于Sepharose 2B)进行压柱,直至凝胶高度恒定。
15.  Mark on the column the position of the bed surface, stop the pump, close the column outlet and adjust the adaptor to the bed surface and then push the adaptor a further 3-4mm.在柱上标记床表面的位置,停止泵压,关闭柱的出口,调整接头至床表面,然后将接头进一步推进3-4mm.

The column is now ready to use.现在柱已备好可用。

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