Montage抗体纯化试剂盒
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概述
规格
订货信息
Documentation
参考
| 参考概述 |
|---|
| Antibody Purification with Affinity Spin Columns Kavonian, Mark,BioScience Technology, July 2003 BioScience Technology, July 2003 2003 |
| Considerations During Development of a Protein A-Based Antibody Purification Process Iyer et al.,Biopharm, January 2002, pp 14-20 Biopharm, January 2002, pp 14-20 2002 |
| A sensitive enzyme linked immunoadsorbent assay (ELISA) for the detection of staphylococcal prtein A (SpA) present as a trace contaminant of murine immunoglobulins purified on immobilized protein A Miguel A.J. Godfrey, Piotr Kwasowski, Roland Clift and Vincent Marks Journal of Immunological Methods, 149(1992) 21-27, 1992 1992 |
问与答
| 问题 | 回答 |
|---|---|
| What is the shelf life of the Prosep A and Prosep G spin columns? | The spin columns/media are under warranty for 2 years provided they are stored at 2-8 C. |
| Do I need to filter the buffers provided with the Montage Antibody Purification kit? | The buffers supplied with the kit are pre-filtered for immediate use. |
| Do I need to pre-filter my sample before loading it onto a Prosep A or ProSep G spin column? | All samples must be filtered through a 0.22 mm filter immediately before loading the sample onto the column. We recommend using a Steriflip-GP 0.22 mm filter. |
| What is the binding capacity of the Prosep A spin column? | Using the Prosep A spin columns we have been able to bind >20 mg of Rabbit IgG. |
| What is the maximum volume I can load onto the Prosep A and Prosep G spin columns? | You can load up to 20 ml in a Prosep A or Prosep G sping column for a swinging bucket rotor and up to 10 ml for a fixed angle rotor. We recommend that you spin it in a swinging bucket rotor. |
| Why do you suggest that the Prosep A spin columns be spun at 100-150 xg for 20 minutes? | The low speed, twenty minute centrifugation maximizes IgG / resin contact which increases IgG binding efficiency. |
| Can I use a fixed angle rotor or a swinging bucket rotor with the Prosep A and Prosep G spin columns? | Swing bucket rotors are recommended because they provide uniform flow through the resin. For optimal performance with a fixed angle rotor, ensure that the orientation of the Prosep A spin coulmn is the same for all steps. Also please note that the maximum sample loading volume for a fixed angle rotor is ten ml versus twenty ml with a swinging bucket rotor. |
| What is the minimum elution volume for the Prosep A and Prosep G spin columns? | We suggest 10 ml. Smaller volumes can be used but they may result in incomplete IgG recovery. If you use a smaller volume the neutralization buffer volume should also be adjusted accordingly. |
| How do I regenerate the Prosep A spin columns? | We recommend that you wash the plugs with 10 ml of Elution Buffer B2 by centrifuging at 500 xg got 5 minutes. re-equilibrate the media with 5 ml of Binding Buffer A by centrifuging the spin column at 500 xg for 2 minutes. Proceed to step 1 of the antibody purification protocol for immediate re-use or store the plugs without their end caps in 5 ml of Binding Buffer a in a 15 ml screw-capped tube for future use. |
| Can I autoclave the Prosep A and Prosep G spin columns? | The Prosep A and Prosep G spin columns cannot be autoclaved. |
相关产品和应用
产品族
Steriflip® 过滤装置不需样品转移，只需把离心柱和过滤器连接起来即可。详细了解 >> |
Amicon Ultra-15离心过滤器提供超快旋转速度和超高回收率两种机型详细了解 >> |
Immobilon® 滤膜、三明治和印迹过滤纸Immobilon P滤膜是适用于西方蛋白印迹检测的理想PVDF转印膜。详细了解 >> |
相关产品，按下列排序： Application Facete
| Protein Purification |
| Dialysis and Filtration |
相关产品，按下列排序： Brand Facete
| Montage® |
| ProSep® |
种类
Montage抗体纯化试剂盒可简单、快捷地从血清、腹水或细胞培养上清液中纯化抗体。试剂盒包括预装了高亲和性的PROSEP-A或G培养基的离心柱。无需安装任何泵， 无需填充任何柱。您只需要用离心机离心不到60分钟，即可纯化10–20 mg。
与传统的重力流方法不同，该实验方案采用离心纯化技术，简单易行，且不需有人在场。
使用结果表明，离心柱可重复使用多达10次，且不损失结合能力或特异性。
性能
离心柱的重复使用
)
使用再生的 PROSEP-G 离心柱从正常的血清人体 IgG 10 次。
MAbs 纯化
)
使用 PROSEP-A 离心柱从腹水对鼠单克隆抗体进行了纯化。 （条带 1：未纯化腹水。 条带 2：渗流的腹水。 条带 3：洗涤渗流的腹水。 条带 4：5µg 纯化的抗体。）承蒙 Exalpha Biologicals 提供。
| 蛋白A和蛋白G的结合亲和性 | ||||||
| 蛋白质 | A | G | 蛋白质 | A | G | |
| 人IgG1 | ++ | ++ | 大鼠IgM | + /-- | - | |
| 人IgG2 | ++ | ++ | 兔IgG | ++ | ++ | |
| 人IgG3 | -- | ++ | 仓鼠IgG | + | ++ | |
| 人IgG4 | ++ | ++ | 豚鼠IgG | ++ | + | |
| 人IgA | + | -- | 牛IgG | + | + | |
| 人IgD | + | -- | 绵羊IgG | + / -- | + | |
| 人IgE | + | -- | 山羊IgG | + /-- | + | |
| 人IgM | + | -- | 猪IgG | ++ | ++ | |
| 小鼠IgG1 | + | + | 鸡IgG | -- | + /-- | |
| 小鼠IgG2a | ++ | ++ | 片段 | |||
| 小鼠IgG2b | ++ | ++ | 人Fab | + | + | |
| 小鼠IgG3 | + | ++ | 人F(ab')2 | + | + /-- | |
| 小鼠IgM | + /-- | -- | 人ScFv | + | -- | |
| 大鼠IgG | ++ | ++ | 人Fc | + | -- | |
| 大鼠IgG1 | + /-- | + | 人K | -- | -- | |
| 大鼠IgG2a | + /-- | ++ | 人λ | -- | + | |
| 大鼠IgG2b | + /-- | + | ||||
| 大鼠IgG2c | + /-- | + | ||||
| 图例：++ 表示强亲和性；+ 表示中等/微弱亲和性；+/-- 表示要求评估；-- 表示无亲和性 | ||||||
| 试剂盒内容 | 包括离心柱、缓冲液、样品制备和浓缩装置 | |
| 支撑基质材料 | 多孔玻璃 | |
| 培养基粒径 | 75 µm–125 µm | |
| 培养基柱床容积 | 1.6 mL | |
| 结合能力 | ||
| 蛋白质A | >20 mg兔IgG | |
| 蛋白质G | >10 mg兔IgG | |
| pH 范围 | pH2 – pH9 | |
| 最大体积 | 20 mL (maximum in swinging bucket rotor) | |
| 保存温度 | ||
| 端盖色码 | ||
| 蛋白质A | 红色 | |
| 蛋白质G | 黄色 | |