重组蛋白定制服务

使用说明书

仅供体外研究使用，不用于临床诊断!

第1版（2016年04月修订）

关联信息

武汉云克隆为您提供从基因到蛋白的一站式服务，客户也可以根据自身的需求灵活的选择服务项目或个性化的定制服务。目前建设拥有原核（E.coli）表达系统与真核（酵母、杆状病毒一昆虫细胞、哺乳动物细胞）表达系统的重组蛋白表达平台。我们将在不断创新和改进的基础上，始终致力于为每一位客户提供最全面、最优质的服务。

表达系统及相关技术优势

杆状病毒-昆虫细胞表达系统

1.多种纯化标签供选择

2. 高滴度病毒制作技术

3. 高效V系列表达载体与快速纯化技术

大肠杆菌表达系统

1. 多种纯化标签供选择

2. 多种表达载体和宿主菌的选择

3. 高效C系列表达载体与快速纯化技术

酵母表达系统

1. 多种纯化标签供选择

2. 多拷贝阳性菌株快速筛选技术

3. 高效Y系列表达载体与快速纯化技术

哺乳动物细胞表达系统

1. 多种纯化标签供选择

2. 哺乳动物细胞高效侵染技术

3. 高效P系列表达载体与快速纯化技术

蛋白纯化

1. 多种纯化方式的选择

2. 快速去内毒素技术

3. 柱上标签切除技术

4. 一步高效纯化技术

5. 包涵体复性

6. EASYFOLD技术

常规流程

基因克隆或者基因合成 →亚克隆质粒构建 →表达质粒构建 →表达条件优化 →蛋白表达 →蛋白纯化 →后期处理：去除内毒素、融合标签切除、活性测定等。

重组蛋白表达服务类型

原核（E.coli）重组蛋白表达服务
真核（酵母）重组蛋白表达服务
真核（杆状病毒-昆虫细胞）重组蛋白表达服务
真核（哺乳动物细胞）重组蛋白表达服务
GMP级别重组蛋白表达服务

GMP级别蛋白实例展示

IS014-Eukaryotic Expressed Human Activin A

Organism Species: Homo sapiens (Human)

PROPERTIES

Source: Eukaryotic expression.

Host: 293F cell 

Residues: Ser21~Ser426

Tags: N-terminal His-Tag

Subcellular Location: Secreted.

Purity: >95%

Endotoxin Level: <1.0EU per 1µg (determined by the LAL method).

Traits: Freeze-dried powder

Buffer formulation: PBS, pH7.4, containing 5% Trehalose .

Original Concentration: 400µg/mL

Applications: Positive Control; Immunogen; SDS-PAGE; WB.

(May be suitable for use in other assays to be determined by the end user.)

Predicted isoelectric point: 7.9

Predicted Molecular Mass: 46.6kDa 

Accurate Molecular Mass: 60&44&13kDa as determined by SDS-PAGE reducing conditions.

Phenomenon explanation:

The possible reasons that the actual band size differs from the predicted are as follows:

1. Splice variants: Alternative splicing may create different sized proteins from the same gene.

2. Relative charge: The composition of amino acids may affects the charge of the protein.

3. Post-translational modification: Phosphorylation, glycosylation, methylation etc.

4. Post-translation cleavage: Many proteins are synthesized as pro-proteins, and then cleaved to give the active form.

5. Polymerization of the target protein: Dimerization, multimerization etc.

USAGE

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE AND STABILITY  

Storage: Avoid repeated freeze/thaw cycles.

               Store at 2-8°C for one month. 

               Aliquot and store at -80°C for 12 months. 

Stability Test: The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

SEQUENCE 

IDENTIFICATION

Figure 1. Gene Sequencing (Extract)

Figure 2. SDS-PAGE

IMPORTANT NOTE

The kit is designed for research use only, we will not be responsible for any issue if the kit was used in clinical diagnostic or any other procedures.

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IS014-Prokaryotic Expressed Human EGF

Organism Species: Homo sapiens (Human)

PROPERTIES

Source: Prokaryotic expression.

Host: E. coli 

Residues: Asn971~Arg1023 

Tags: N-terminal His-Tag

Subcellular Location: Membrane.

Purity: >98%

Endotoxin Level: <1.0EU per 1µg (determined by the LAL method).

Traits: Freeze-dried powder

Buffer formulation: PBS, pH7.4, containing 0.01% SKL, 5% Trehalose .

Original Concentration: 100µg/mL

Applications: Positive Control; Immunogen; SDS-PAGE; WB.

(May be suitable for use in other assays to be determined by the end user.)

Predicted isoelectric point: 4.8

Predicted Molecular Mass: 9.9kDa 

Accurate Molecular Mass: 10kDa as determined by SDS-PAGE reducing conditions.

USAGE

Reconstitute in ddH2O to a concentration of 0.1-0.5 mg/mL. Do not vortex.

STORAGE AND STABILITY  

Storage: Avoid repeated freeze/thaw cycles.

               Store at 2-8°C for one month. 

               Aliquot and store at -80°C for 12 months. 

Stability Test: The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

SEQUENCE 

IDENTIFICATION

Figure 1. Gene Sequencing (Extract)

Figure 2. SDS-PAGE

IMPORTANT NOTE

The kit is designed for research use only, we will not be responsible for any issue if the kit was used in clinical diagnostic or any other procedures.

---

IS014-Eukaryotic Expressed Human FGF7

Organism Species: Homo sapiens (Human)

PROPERTIES

Source: Eukaryotic expression.

Host: 293F cell 

Residues: Cys32~Thr194

Tags: N-terminal His-Tag

Subcellular Location: Secreted.

Purity: >95%

Endotoxin Level: <1.0EU per 1µg (determined by the LAL method).

Traits: Freeze-dried powder

Buffer formulation: PBS, pH7.4, containing 5% Trehalose.

Original Concentration: 80µg/mL

Applications: Positive Control; Immunogen; SDS-PAGE; WB.

(May be suitable for use in other assays to be determined by the end user.)

Predicted isoelectric point: 9.5

Predicted Molecular Mass: 19.4kDa 

Accurate Molecular Mass: 25kDa as determined by SDS-PAGE reducing conditions.

Phenomenon explanation:

The possible reasons that the actual band size differs from the predicted are as follows:

1. Splice variants: Alternative splicing may create different sized proteins from the same gene.

2. Relative charge: The composition of amino acids may affects the charge of the protein.

3. Post-translational modification: Phosphorylation, glycosylation, methylation etc.

4. Post-translation cleavage: Many proteins are synthesized as pro-proteins, and then cleaved to give the active form.

5. Polymerization of the target protein: Dimerization, multimerization etc.

USAGE

Reconstitute in ddH2O to a concentration ≤0.1mg/mL. Do not vortex.

STORAGE AND STABILITY  

Storage: Avoid repeated freeze/thaw cycles.

               Store at 2-8°C for one month. 

               Aliquot and store at -80°C for 12 months. 

Stability Test: The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

SEQUENCE 

IDENTIFICATION

Figure 1. Gene Sequencing (Extract)

Figure 2. SDS-PAGE

IMPORTANT NOTE

The kit is designed for research use only, we will not be responsible for any issue if the kit was used in clinical diagnostic or any other procedures.