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Anti-EGFRvIII Monoclonal Antibody, recombinant

Catalog #: EG-369


$250.00

Qty:

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 Ⅲ (EGFRvⅢ) 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). EGFRⅢ expression has not been found in normal tissues [2]. Expression of EGFRvIII is considered a negative prognostic indicator. These attributes make EGFRⅢ 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
EGFRvIII-R6-flow.jpg
 
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.

     
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