分子别名(Synonym)
EGFRvIII
表达区间及表达系统(Source)
Alexa Fluor 488-Labeled Human EGFRvIII, His Tag (EGI-HA2H7) is produced via conjugation of AF488 to Human EGFRvIII, His Tag with a new generation site-specific technology under Star Staining labeling platform. Human EGFRvIII, His Tag is expressed from human 293 cells (HEK293). It contains AA Leu 25 - Ser 378 (Accession # NP_001333870.1).
Predicted N-terminus: Leu 25
蛋白结构(Molecular Characterization)
This protein carries a polyhistidine tag at the C-terminus
The protein has a calculated MW of 53.0 kDa.
偶联(Conjugate)
AF488
Excitation Wavelength: 488 nm
Emission Wavelength: 517 nm
内毒素(Endotoxin)
Less than 1.0 EU per μg by the LAL method.
纯度(Purity)
>90% as determined by SDS-PAGE.
制剂(Formulation)
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4 with trehalose as protectant.
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 protect from light and avoid repeated freeze-thaw cycles.
This product is stable after storage at:
- -20°C to -70°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.
CAR阳性表达率检测(Evaluation of CAR expression)
FACS Analysis of Anti-EGFRvIII CAR Expression
5e5 of anti-EGFRvIII CAR-293 cells were stained with 100 μL of 20 μg/mL of Alexa Fluor 488-Labeled Human EGFRvIII, His Tag (Cat. No. EGI-HA2H7) and negative control protein respectively (Fig. C and B), and non-transfected 293 cells were used as a control (Fig. A). Alexa Fluor 488 signal was used to evaluate the binding activity (QC tested).
Protocol
FACS Analysis of Non-specific binding to PBMCs
5e5 of PBMCs were stained with Alexa Fluor 488-Labeled Human EGFRvIII, His Tag (Cat. No. EGI-HA2H7) and anti-CD3 antibody, washed and then analyzed with FACS. PE signal was used to evaluate the expression of CD3+ T cells in PBMCs, and Alexa Fluor 488 signal was used to evaluate the non-specific binding activity to PBMCs (QC tested).
Protocol
背景(Background)
The epidermal growth factor receptor (EGFR; ErbB-1; HER1 in humans) is the cell-surface receptor for members of the epidermal growth factor family (EGF-family) of extracellular protein ligands. The epidermal growth factor receptor is a member of the ErbB family of receptors, a subfamily of four closely related receptor tyrosine kinases: EGFR (ErbB-1), HER2/c-neu (ErbB-2), Her 3 (ErbB-3) and Her 4 (ErbB-4). Mutations affecting EGFR expression or activity could result in cancer.
The type III EGF deletion mutant receptor (EGFRvIII) is the most common mutation and was first identified in primary human glioblastoma tumors. This tumor-specific antigen is ligand-independent, contains a constitutively active tyrosine kinase domain, and has been shown to be present in a number of human malignancies. EGFRvIII has been selected as a target for CAR-modified T-cell studies in recent years.