His Pull-Down 磁珠

产品名称：Ni charged Magnetic Beads for His Pull-Down Assay， His Pull-Down 磁珠

货号： IF9301

品牌：Engibody

简介：His Pull-Down 磁珠，即镍磁珠，用于Pull-Down实验中抓取his融合蛋白与互作蛋白的复合物

• Discover a new protein:protein interaction from a cell lysate

• Confirm a putative interaction from a cell lysate or with a previously purified protein

• Extract protein:protein interaction information from in vitro transcription/translation lysates

Tested applications

His Pull-Down Assay.

Optimal amount should be determined by the end user.

Specificity

Binding His-Tagged Fusion Protein.

Form

Magnetic Beads in Suspension (20% 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, dry or centrifuge magnetic beads. This may cause irreversible aggregation and decreased binding capacity.

Bead Diameter:40 µm

Concentration:20% slurry

Binding Capacity:5 to10 mg His fusion protein per 1 mL settled beads (5mL 20% slurry).

Protocols

Procedure Summary for His Pull-Down Assay

I. Preparation of Bait Protein  

Protocol 1: Bait from E. coli Expression Systems

Protocol 2: Bait from Insect Cell Expression Systems

Protocol 3: Bait from Previously Purified Protein

II. Immobilization of Bait Protein

A. Preparation of Glutathione beads

B. Immobilize Bait Protein

III. Preparation of Prey Protein

Protocol 1: Prey Protein from E. coli or Baculovirus-Infected Insect Cell Lysate

Protocol 2: Prey Protein from Mammalian Cell Lysate

Protocol 3: Prey from Previously Purified Protein

Protocol 4: Prey Protein from In vitro Transcription/Translation Reaction

IV. Capture of Prey Protein

V. Elution of Bait-Prey

VI. SDS PAGE Gel Analysis or WB Analysis

用途

•发现一种新的蛋白质：来自细胞裂解物的蛋白质相互作用

•确认来自细胞裂解物或与先前纯化的蛋白质的假定相互作用

•提取蛋白质：来自体外转录/翻译裂解物的蛋白质相互作用信息

经过测试的应用

His蛋白的相互作用蛋白的pull down试验。

最佳用量应由最终用户确定。

特异性

结合His标记的融合蛋白。

类型

悬浮液中的磁珠（20%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。请勿冷冻、干燥或离心磁珠。这可能导致不可逆聚集和结合能力降低。

磁珠直径：40µm

浓度：20%浆料

结合能力：每1mL沉淀磁珠（5mL 20%浆料）5至10mg His融合蛋白。

操作流程

一、 诱饵蛋白的制备

方案1：来自大肠杆菌表达系统的诱饵

方案2：来自昆虫细胞表达系统的诱饵

方案3：来自先前纯化蛋白的诱饵

二、诱饵蛋白的固定化

A、 GSH磁珠的准备

B、 固定化诱饵蛋白

三、 猎物蛋白的制备

方案1：来自大肠杆菌或杆状病毒感染的昆虫细胞裂解物的捕食蛋白

方案2：来自哺乳动物细胞裂解物的猎物蛋白

方案3：来自先前纯化的蛋白质的猎物

方案4：来自体外转录/翻译反应的猎物蛋白

四、 捕获猎物蛋白质

五、 饵饵的洗脱

六、 SDS PAGE凝胶分析或WB分析