United Kingdom (UK)
Recombinant monoclonal antibody to EGFRvIII. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma L8A4.
UniProt Accession Number of Target Protein: P00533
Alternative Name(s) of Target: Epidermal Growth Factor Receptor variant III
Immunogen: Pep 3-KLH (Pep 3 is a 14-amino acid peptide corresponding to the predicted amino acid sequence at the fusion junction and KLH stands for keyhole limpet hemocyanin) at da 40 and 103, HC2 20 d2 cells at day 1, 25, 74, 87. And fusion with the nonimmunoglobulin-secreting Kearney variant of P3X63/Ag8.653.
Specificity: Binds EGFRvIII, a mutated form of EGFR, plays a prominent role in tumorigenesis.
Application Notes: This antibody can be used to detect EGFRvIII, a mutated EGFR variant, in immunohistochemical staining, Western Blot, ELISA and flow-cytometry. Chou et al (2016, PMID: 27825958) used this antibody to detect EGFRvIII in acetone and PFA fixed sections by fluorescent immunohistochemistry.
Antibody first published in:
Wikstrand CJ, Hale LP, Batra SK, Hill ML, Humphrey PA, Kurpad SN, McLendon RE, Moscatello D, Pegram CN, Reist CJ, et al. Monoclonal antibodies against EGFRvIII are tumor specific and react with breast and lung carcinomas and malignant gliomas. Cancer Res. 1995 Jul 15;55(14):3140-8. PMID:7606735
Note on publication:
Describes the preparation anti human EGFRvIII antibody by long-termimmunization protocols using the EGFRvIII-specific synthetic peptide and the intact variant. Identified using ELISA, Western blot, flow cytometry, and immunohistochemistry techniques.
Flow-cytometry using the anti-EGFRvIII antibody L8A4 (Ab00184) A431 cells were fixed using 2% PFA, permeabilised using 0.5% Triton and stained with anti-Fluorescein IgG antibody (4-4-20; isotype control, black line) or the mouse IgG1 version of L8A4 (Ab00184-1.1, blue line) at a dilution of 1:100 for 1h at RT. After washing, bound antibody was detected using a goat anti-mouse IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.