United Kingdom (UK)
Recombinant monoclonal antibody to EGFR. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the scFv L3 11D.
UniProt Accession Number of Target Protein: P00533
Alternative Name(s) of Target: Epidermal growth factor receptor; ERBB; ERBB1; HER1; mENA; PIG61; NISBD2
Immunogen: Human EGFR.
Specificity: The antibody binds to EGFR with an affinity (Kd) of 0.21 nM.
Application Notes: The antibody binds specifically to EGFR, a cell-surface receptor which is part of the ErbB family of receptors. The EGFR is a receptor tyrosine kinase, which, upon activation by its ligands, such as epidermal growth factor and transforming growth factor alpha, triggers signal transduction pathways that increase cell migration, adhesion and proliferation. Mutations that lead to the overexpression of EGFR are associated with the development of cancer. The antibody was prepared from phage antibodies isolated from the lymph node library.
Antibody first published in:
Kettleborough et al. Isolation of tumor cell-specific single-chain Fv from immunized mice using phage-antibody libraries and the re-construction of whole antibodies from these antibody fragments. Eur. J. Immunol. 24 (4), 952-958 (1994) PMID:8149964
Note on publication:
Describes the phage-antibody technology, which is used to produce three phage-antibody libraries from which mouse anti-EGFR scFv antibodies are generated. The libraries are prepared from the spleen, lymph node and in vitro cells of an immunized mouse. The antibodies produced are compared with a mouse mAb, 425, isolated by standard hybridoma technology.
Immunofluorescence staining of A431 cells using anti-EGFR antibody (Ab00455) L3 11D. Immunofluorescence analysis of paraformaldehyde fixed A431 cells stained with the chimeric rabbit IgG version of L3 11D (Ab00455-23.0) at 10 µg/ml followed by Alexa Fluor® 488 secondary antibody (2 µg/ml), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab00455-23.0, DAPI, merged channels and an isotype control. The isotype control was stained with anti-Fluorescein antibody (Ab00102-23.0) followed by Alexa Fluor® 488 secondary antibody.
Flow-cytometry using the anti-EGFR antibody L3 11D (Ab00455). A431 cells were stained with anti-Fluorescein IgG antibody (4-4-20; isotype control, black line) or the rabbit IgG1 version of L3 11D (Ab00455-23.0, 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.
Immunofluorescence staining of A431 cells using anti-EGFR antibody (Ab00455) L3 11D. Immunofluorescence analysis of paraformaldehyde fixed A431 cells stained with the chimeric mouse IgG version of L3 11D (Ab00455-1.1) at 10 µg/ml followed by Alexa Fluor® 488 secondary antibody (2 µg/ml), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab00455-1.1, DAPI, merged channels and an isotype control. The isotype control was stained with anti-unknown antibody (Ab00178-1.1) followed by Alexa Fluor® 488 secondary antibody.
Flow-cytometry using the anti-EGFR antibody L3 11D (Ab00455). A431 cells were stained with anti-unknown IgG antibody (Ab00178-1.1; isotype control, black line) or the mouse IgG-chimeric version of L3 11D (Ab00455-1.1, 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.
Flow cytometry using the Anti-EGFR antibody L3 11D (Ab00455). A431 cells were stained with anti-unknown specificity antibody (Ab00178-1.1; isotype control, black line) or the mouse IgG1 version of L3 11D (Ab00455-1.1, blue line) at a dilution of 1:100 for 1h at RT. After washing, the 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.