Streptavidin琼脂糖珠

产品名称：Streptavidin Agarose Beads， Streptavidin琼脂糖珠

货号： IF9041

品牌：Engibody

关键词：Streptavidin琼脂糖珠，用于蛋白质、抗体、肽和其他biotin化分子的偶联。

Streptavidin is a 53,000 dalton tetrameric protein purified from the bacterium Streptomyces avidinii. Each subunit binds to biotin with extremely high affinity. Because of its strong non-covalent interaction with biotin, streptavidin can be used to isolate biotinylated proteins.

Description

Streptavidin Agarose Beads is useful for the precipitation of biotinylated proteins. Recombinant streptavidin is immobilized via covalent binding of primary amino groups to N-hydroxysuccinimide (NHS)-activated agarose beads.

Tested applications

For coupling of proteins, antibodies, peptides and other molecules. Optimal amount should be determined by the end user.

Form

Beads in suspension (50% slurry), supplied in phosphate buffered saline (PBS) pH 7.4, with 0.02% sodium azide as preservatives.

Storage instruction

Store at 4°C. This product is stable for 12 months. Do not freeze.

Beads Diameter

90 µm

Binding Capacity

>100nmol Free biotin

200μg Biotinylated antibody(/mL beads slurry)

Protocols

Additional information

Streptavidin Agarose Beads should be resuspended well before used.

Beads cannot be reused.

Procedure for binding biotinylated target directly.

A. Solutions and Reagents

NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.

1. Recommended coupling/washing buffers

protein/antibody samples: PBS buffer, pH7.4, nucleic acids samples: TES buffer

2. Elution buffer (only for biotinylated target purification): 8M Guanidine hydrochloride, pH1.5

B. Wash Streptavidin agarose beads

Calculate the amount of beads required based on their binding capacity (see the above Binding Capacity), and transfer the beads to a new tube.

1. Resuspend the beads in the vial (i.e. tilt and rotate for 5 min).

2. Transfer the desired volume of beads to a tube.

3. Add an equal volume of Washing buffer, or at least 1 mL, and mix (keep on a roller for at least 5 min).

4. Centrifuge the tube at 4000 g for 1 min and discard the supernatant.

5. Resuspend the washed beads in the same volume of washing buffer as the initial volume of beads taken from the vial (step 2).

C. Binding protein/antibody

1. Incubate the beads and biotinylated protein/antibody in PBS buffer (pH 7.4) for 30 min at room temperature using gentle rotation.

2. Centrifuge the protein/antibody-coated beads at 4000 g for 2–3 min and discard the supernatant.

3. Wash the coated beads 4–5 times in PBS containing 0.1% BSA.

4. Resuspend to the desired concentration for your application.

NOTE:

Elution of the biotinylated proteins that are bound to the Streptavidin Beads requires harsh conditions, such as boiling for 5 minutes with 8M guanidine•HCl, pH 1.5. Protein may leach from the beads with such conditions after centrifugation.

链霉亲和素是一种53000道尔顿的四聚体蛋白，从细菌阿维迪尼链霉菌中纯化。每个亚基以非常高的亲和力与biotin结合。由于其与biotin的强非共价相互作用，链霉亲和素可用于分离biotin化蛋白。

描述

链霉亲和素琼脂糖珠可用于biotin化蛋白的沉淀。重组链霉亲和素通过伯氨基与N-羟基琥珀酰亚胺（NHS）活化的琼脂糖珠共价结合而固定。

经过测试的应用

用于蛋白质、抗体、肽和其他biotin化分子的偶联。最佳用量应由最终用户确定。

类型

悬浮珠（50%浆料），在pH 7.4的磷酸盐缓冲盐水（PBS）中提供，含0.02%防腐剂。

存储说明

储存于4°C。该产品稳定12个月。不要冻结。

beads直径

90µm

结合能力

>100nmol游离biotin

200μg的biotin化抗体（/mL珠浆）

链霉亲和素琼脂糖珠应在使用前重新悬浮。

珠子不能重复使用。

操作流程

直接结合biotin化靶分子的流程

A、 溶液和试剂

注：用反渗透去离子水（RODI）或同等级别的水制备溶液。

1.推荐的偶联和清洗缓冲液

蛋白质/抗体样品：PBS缓冲液，pH7.4，核酸样品：TES缓冲液

2.洗脱缓冲液（仅用于biotin化目标纯化）：8M盐酸胍，pH1.5

B、 清洗链霉亲和素琼脂糖珠

根据珠子的结合能力计算所需的珠子数量（参见上述结合能力），并将珠子转移到新的试管中。

1.将珠子重新悬浮在小瓶中（即倾斜并旋转5分钟）。

2.将所需体积的珠子转移到试管中。

3.加入等量的洗涤缓冲液，或至少1mL，并混合（在滚筒上保持至少5分钟）。

4.以4000g离心试管1分钟，并丢弃上清液。

5.将洗涤过的珠粒重新悬浮在与从小瓶中取出的珠粒的初始体积相同体积的洗涤缓冲液中（步骤2）。

C、 结合蛋白/抗体

1.将珠粒和biotin化蛋白/抗体在PBS缓冲液（pH 7.4）中在室温下使用温和旋转孵育30分钟。

2.以4000 g离心蛋白/抗体包被的珠粒2-3分钟，并丢弃上清液。

3.在含有0.1%BSA的PBS中清洗涂覆的珠粒4-5次。

4.重新悬浮至您所需的浓度。

注意：

与链霉亲和素珠结合的biotin化蛋白质的洗脱需要苛刻的条件，例如用8M胍•HCl，pH 1.5煮沸5分钟。离心后，蛋白质可能在这种条件下从珠中浸出。