重組蛋白定制服務(wù)

使用說明書

僅供體外研究使用，不用于臨床診斷!

第1版（2016年04月修訂）

關(guān)聯(lián)信息

武漢云克隆為您提供從基因到蛋白的一站式服務(wù)，客戶也可以根據(jù)自身的需求靈活的選擇服務(wù)項(xiàng)目或個(gè)性化的定制服務(wù)。目前建設(shè)擁有原核（E.coli）表達(dá)系統(tǒng)與真核（酵母、桿狀病毒一昆蟲細(xì)胞、哺乳動(dòng)物細(xì)胞）表達(dá)系統(tǒng)的重組蛋白表達(dá)平臺(tái)。我們將在不斷創(chuàng)新和改進(jìn)的基礎(chǔ)上，始終致力于為每一位客戶提供最全面、最優(yōu)質(zhì)的服務(wù)。

表達(dá)系統(tǒng)及相關(guān)技術(shù)優(yōu)勢(shì)

桿狀病毒-昆蟲細(xì)胞表達(dá)系統(tǒng)

1.多種純化標(biāo)簽供選擇

2. 高滴度病毒制作技術(shù)

3. 高效V系列表達(dá)載體與快速純化技術(shù)

大腸桿菌表達(dá)系統(tǒng)

1. 多種純化標(biāo)簽供選擇

2. 多種表達(dá)載體和宿主菌的選擇

3. 高效C系列表達(dá)載體與快速純化技術(shù)

酵母表達(dá)系統(tǒng)

1. 多種純化標(biāo)簽供選擇

2. 多拷貝陽(yáng)性菌株快速篩選技術(shù)

3. 高效Y系列表達(dá)載體與快速純化技術(shù)

哺乳動(dòng)物細(xì)胞表達(dá)系統(tǒng)

1. 多種純化標(biāo)簽供選擇

2. 哺乳動(dòng)物細(xì)胞高效侵染技術(shù)

3. 高效P系列表達(dá)載體與快速純化技術(shù)

蛋白純化

1. 多種純化方式的選擇

2. 快速去內(nèi)毒素技術(shù)

3. 柱上標(biāo)簽切除技術(shù)

4. 一步高效純化技術(shù)

5. 包涵體復(fù)性

6. EASYFOLD技術(shù)

常規(guī)流程

基因克隆或者基因合成 →亞克隆質(zhì)粒構(gòu)建 →表達(dá)質(zhì)粒構(gòu)建 →表達(dá)條件優(yōu)化 →蛋白表達(dá) →蛋白純化 →后期處理：去除內(nèi)毒素、融合標(biāo)簽切除、活性測(cè)定等。

重組蛋白表達(dá)服務(wù)類型

原核（E.coli）重組蛋白表達(dá)服務(wù)
真核（酵母）重組蛋白表達(dá)服務(wù)
真核（桿狀病毒-昆蟲細(xì)胞）重組蛋白表達(dá)服務(wù)
真核（哺乳動(dòng)物細(xì)胞）重組蛋白表達(dá)服務(wù)
GMP級(jí)別重組蛋白表達(dá)服務(wù)

GMP級(jí)別蛋白實(shí)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.

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.