免疫亲和色谱柱制备方法

Instructions 71-7086-00 AF Affinity media

GE Healthcare CNBr-activated Sepharose™ 4B CNBr-activated Sepharose 4B is a pre-activated medium for immobilization of ligands containing primary amines. It provides a very convenient way to immobilize ligands by the cyanogen bromide method. The coupling reaction is spontaneous, rapid and easy to carry out. The application area covers immobilization of proteins, peptides and nucleic acids.

通用电气医疗集团CNBr-activated琼脂糖™4 b CNBr-activated琼脂糖4 b是固定地激活介质的包含一级胺配体。它提供了一个非常方便的方法固定溴化氰配体的方法。偶联反应是自发的,快速和容易执行。应用领域涵盖固定的蛋白质、多肽和核酸。

Table 1. Medium characteristics.

Bead structure: 4% agarose

Spacer: None

Coupling capacity: 25–60 m g α-chymotrypsinogen/ml drained medium Bead size range: 45–165 μm

Average bead size: 90 μm

Max linear flow rate*: 75 cm/h at 25°C, HR 16/10 column, 5 cm bed height

pH stability**:

表1。媒介的特征。

琼脂糖珠结构:4%

间隔:没有

耦合能力:α-chymotrypsinogen外地毫克/毫升排水

媒介

珠大小范围:45 - 165μm

平均珠大小:90μm

最大线性流量*:75厘米/小时25°C,人力资源列16/10,5厘米床层高度pH值稳定* *:

Long term: 3–11

Short term: 2–11

Chemical stability***: Stable to all commonly used aqueous solutions. Can be used with non-ionic detergents, denaturing

solvents, e.g. 8 M urea and 6 M guanidine hydrochloride. Stable in organic solvents, such as 50% dimethylformamide and 50% dioxane. Autoclavable: Not recommended

长期:3-11

短期:2 - 11

化学稳定性* * *:所有常用的水稳定的解决方案。

可以用于非离子去污剂,变性

溶剂,例如8 M尿素和6 M胍盐酸。等有机溶剂、稳定50%二甲基甲酰胺和二氧六环的50%。

Autoclavable:不推荐

* Linear flw rate = volumetric flw rate (cm 3/h) /column cross-sectional

area (cm 2)

** The ranges given are estimates based on our knowledge and experience. Please note the following:

pH stability, long term refers to the pH interval where the gel is stable over a long period of time without adverse effects on its subsequent chromatographic performance.

*线性flw率=容积flw率(厘米3 /小时)

列横截面积(2厘米)

* *给出的范围的估计是基于我们的知识和经验。请

请注意以下几点:

pH值稳定、长期的pH值间隔指的是凝胶稳定在很长一段时间没有不利影响在其随后的色谱性能。

pH stability, short term refers to the pH interval for regeneration, cleaning-in-place and sanitization procedures.

*** Data refer to the coupled product, provided that the ligand can withstand the pH or chemical environment.

pH值稳定,短期内指的是再生的pH值间隔,原位清洗（CIP）和卫生处理程序。

* * *数据指的是耦合的产品,提供的配体能够承受或化学环境pH值。Contents

1. Preparing the medium 4

2. Coupling the ligand 4

3. Factors effecting the coupling efficiency 5

4. Packing Sepharose 4B 7

5. Binding 9

6. Elution 9

7. Regeneration 10

8. Storage 10

9. Further information 10

10. Ordering information 11

内容

1。准备中4

2。耦合配体4

3。因素影响耦合效率的5

4。包装琼脂糖4 b 7

5。绑定9

6。洗脱9

7。再生10

8。存储10

9。进一步的信息10

10。订购信息11

1. Preparing the medium

CNBr-activated Sepharose 4B is supplied lyophilized in the presence of additives. These additives must be washed away at low pH (pH 3) before