这是使用 LudgerClean™ S 多糖纯化柱(Ludger Cat# LC-S-A6)的步骤图文. 这些柱子用于纯化荧光标记(如 2-AB, 2-AA 或普鲁卡因胺标记）后的聚糖。
本文作为 LC-S-A6 产品说明书的完整补充。
链接：http://www.ludgersh.com/9ysh/up/updown/2017112453749781.pdf
This is a step-by-step visual guide to using our LudgerClean S glycan clean up
cartridges (Ludger Cat# LC-S-A6). You would typically use these for purification
of labeled glycans after fluorescent tagging e.g. 2-AB, 2-AA or procainamide.
This document complements the full LC-S-A6 product guide.
1 多糖样品的制备 Prepare the glycan sample
需要纯化的样品体积最多是 15μl。如果您的样品体积较多，那就使用离心蒸发器烘干后重新加不超过
15μl 的水配制。
The sample to be cleaned must have a volume of 15 μl or less. If your sample has a greater volume then dry it down using a
centrifugal evaporator and reconstitute in not more than 15 μl of water
预备 LudgerClean™ S 多糖纯化柱（每样本用一个）的冲洗方式：
• 第一次冲洗 - 1 ml 水冲洗
• 第二次冲洗 - 5 ml 30% 醋酸 (aq) 冲洗
• 第三次冲洗 - 1 ml 乙腈冲洗
让每一次冲洗完全排干净后再添加下一个溶液。如果流量受阻,例如被气隙堵塞, 可以在柱子的顶部施加轻
微的压力,以恢复正常流动。
Ludger Document # LC-S-A6-Photo-Guide-v1.0
© Ludger Limited, 2015 Page 2
2 预备 LudgerClean™ S 多糖纯化柱 Prime the LudgerClean™ S cartridge
Prime the LudgerClean™ S cartridges (one per sample) by washing each with the following :
• 1st wash - 1 ml water
• 2nd wash - 5 ml 30 % acetic acid (aq)
• 3rd wash - 1 ml acetonitrile
Allow each wash to drain completely before adding the next. If flow is restricted, e.g. by an air gap, apply a slight
pressure to the top of the cartridge in order to resume normal flow.
3 在柱子的膜上进行点样 Spot sample onto cartridge membrane
把每个样本点到刚洗过的柱子板上，要确保板仍然被乙腈浸湿。如果可以，在整个板的表面散开点样,因为
这样能帮助清洗。
Spot each sample onto a freshly washed cartridge disc ensuring that the disc is still wet with acetonitrile. Spread the spot over
the entire disc surface if possible as this aids cleanup.
4 让样品吸附到薄膜上 Allow sample to adsorb onto membrane
允许吸附 15 分钟。
Allow adsorption for 15 minutes.
5 从样品瓶中添加剩余样品 Add residual sample from sample vial
用 100μl 乙腈冲洗每个样品瓶然后将冲洗添加到相应的柱子板上。
Rinse each sample vial with 100 μl acetonitrile and apply to the corresponding cartridge disc.
6 洗去膜上的非聚糖杂质 Wash non-glycan contaminants off membrane
用1毫升乙腈冲洗每个板,然后再用 5x1ml 96% 乙腈* / 4% 的水冲洗
*(对于O-聚糖或用普鲁卡因胺标记的N-和O-聚糖的清洗, 用 100% 乙腈做替代)
Wash each disc with 1 ml acetonitrile, followed by 5 x 1 ml 96 % acetonitrile* / 4% water
*(for cleanup of O-glycans or N- and O-glycans labeled with procainamide, substitute with 100% acetonitrile)
7 用 2x0.5ml 水从柱子薄膜上将聚糖洗脱到合适的的容器里。
Elute glycans off membrane into a suitable container by eluting with 2 x 0.5 ml water.
加入下一次 0.5 ml 水之前，确保每 0.5 ml 样本充分流出。
Allow each 0.5 ml aliquot to drain before the next is applied.
8 干燥洗脱下来的聚糖(可选) Dry the eluted glycans (optional)
如果适宜, 蒸发洗脱的聚糖至干燥状态, 然后溶于一定体积的水或溶剂为进行下一步分析。
If appropriate, evaporate the glycan containing fraction to dryness, then dissolve in a desired volume of water or solvent for further analysis.
9 流程完毕 Protocol Complete
你的聚糖已经可以进行分析了。
Your glycans are now ready to analyse.
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