Biotinylated Human EGF R Protein, His,Avitag™ (MALS verified)

Biotinylated Human EGF R Protein, His,Avitag™ (MALS verified)

分子别名（Synonym）

EGFR,ERBB,ERBB1,HER1,PIG61,mENA

表达区间及表达系统（Source）

Biotinylated Human EGF R, His,Avitag is expressed from human 293 cells . It contains AA Leu 25 - Ser 645 

Predicted N-terminus: Leu 25

Request for sequence

蛋白结构（Molecular Characterization）

This protein carries a polyhistidine tag at the C-terminus, followed by an Avi tag (Avitag™).

The protein has a calculated MW of 72.3 kDa. The protein migrates as 90-125 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.

标记（Biotinylation）

Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.

蛋白标记度（Biotin:Protein Ratio)

Passed as determined by the HABA assay / binding ELISA.

内毒素（Endotoxin）

Less than 1.0 EU per μg by the LAL method.

纯度（Purity）

>90% as determined by SDS-PAGE.

>90% as determined by SEC-MALS.

制剂（Formulation）

Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally trehalose is added as protectant before lyophilization.

Contact us for customized product form or formulation.

重构方法（Reconstitution）

Please see Certificate of Analysis for specific instructions.

For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.

存储（Storage）

For long term storage, the product should be stored at lyophilized state at -20°C or lower.

Please avoid repeated freeze-thaw cycles.

This product is stable after storage at:

1. -20°C to -70°C for 24 months in lyophilized state;

2. -70°C for 12 months under sterile conditions after reconstitution.

 

电泳（SDS-PAGE）

Biotinylated Human EGF R, His,Avitag on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 90%.

The purity of Biotinylated Human EGF R, His,Avitagis more than 90% and the molecular weight of this protein is around 100-115 kDa verified by SEC-MALS.

Report

 

活性（Bioactivity）-ELISA

Immobilized Cetuximab at 2 μg/mL (100 μL/well) can bind Biotinylated Human EGF R, His,Avitag with a linear range of 2-6 ng/mL (QC tested).

Protocol

Immobilized Human EGF, Tag Free  at 10 μg/mL (100 μL/well) can bind Biotinylated Human EGF R, His,Avitag  with a linear range of 0.156-5 μg/mL (Routinely tested).

Protocol

CAR阳性表达率检测（Evaluation of CAR expression ）

FACS Analysis of biotinylated EGFR binding cell surface anti-EGFR scFv

Cells were transfected with anti-EGFR scFv and cultured for 3 days. Three days post-transfection, 1e6 cells were stained for anti-EGFR scFv with Biotinylated Human EGFR, His,Avitag  followed by PE-conjugated streptavidin. Flow Cytometry assay shows that Biotinylated Human EGFR, His,Avitag  can bind to cells expressing anti-EGFR scFv. The concentration of EGFR used is 0.8 µg/mL. (Data are kindly provided by Guangzhou Bio-gene Technology Co. Ltd.)

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