| Product Name |
Anti-EGFRvIII monoclonal antibody, recombinant |
| Cat # |
EG-369 |
| Size |
100 µg |
| Key features |
Rabbit monoclonal antibody produced in 293 cells with serum-free, protein-free medium, purified with protein A column;
High batch-to-batch consistency and reproducibility;
Long term security of supply; |
| Clone number |
R6 |
| Immunogen |
Proprietary peptide; |
| Reaction target |
Human EGFRvIII mutant; |
| Species Cross-Reactivity |
Human |
| Applications |
Flow Cytometry; IP; ELISA; Western blot |
| Working concentrations |
Flow cytometry and ELISA: 5 µg/ml of final concentration is suggested.
IP: 10 μg in 0.3-0.5 mg cell lysates is suggested.
Western blot: 1µg/ml of final concentration is suggested. |
| Background |
Epidermal growth factor receptor variant III (EGFRvIII) is the most common EGFR mutation that occurs in up to 30% of high-grade gliomas, especially glioblastoma multiforme (GBM). Epidermal growth factor receptor variant â…¢ (EGFRvâ…¢) is a cancer-specific deletion mutant that is restrictively expressed in glioblastoma and some lung cancers. EGFRvIII plays an important role in tumorigenesis, disease progression, and metastasis, and was identified as a cancer driver mutation gene in glioblastoma patients (1). EGFRâ…¢ expression has not been found in normal tissues [2]. Expression of EGFRvIII is considered a negative prognostic indicator. These attributes make EGFRâ…¢ a perfect target for immunotherapy.
References:
1. Yang J., et al. Cancer Lett. 2017;403:224-30
2. Jungbluth A.A., et al. PNAS. 2003; 100: 639-644 |
| QC |
Fig. A) Flow cytometry assay showing cell binding specificity: one million of U87 cells (EGFRvIII negative), U87 (EGFR++) that was forcefully over-expressing wild type EGFR served as target negative control. U87 (EGFRvIII++) that was forcefully over-expressing EGFRvIII served as test cells. Cells were stained with 5 μg antibody in 1 ml of PBS buffer containing 5% BSA. Antibody binding was detected with Cy5-conjugated goat-anti-rabbit antibody. Shaded area, isotype control antibody staining. Red line, EGFRvIII antibody staining.
Fig.B) ELISA assay showing target protein binding specificity: recombinant EGFR-hFc (serving as negative control), EGFRvIII-hFc (target), and BSA was immobilized on ELISA plate at 5 μg/ml. Biotinylated antibody (1:2 serial dilutions starting from 200 nM) was incubated with the plate at 37°C for 30 min. Antibody binding was visualized with HRP-conjugated streptavidin. |
| Concentrations |
As indicated on the vials |
| Formulation |
80% PBS, pH 7.4, 20% Glycerol, 5% Trehalose, 0.04% Sodium azide as Preservative. |
| Shipping |
Blue ice |
| Storage |
Upon receipt, store it immediately at -20 or -70 °C (long term) or 4 °C (1 week).
Depending on the expected frequency of use, the product can be aliquoted to avoid heavily repeated freezing/thawing cycles. |