Recombinant Mouse Antibody (7A7) is capable of binding to Human EGFR, expressed in Chinese Hamster Ovary cells (CHO). 7A7 recognizes a site on the EGFR extracellular domain that is not accessible in its most stable conformations, but that becomes exposed upon treatment with a tyrosine kinase inhibitor.
Figure 2 7A7 and 7A7 Fc Silent do not detect EGFR expression by flow cytometry.
Human (A431) and mouse (3T3-L1, HPV38 and TC-1) cell lines were stained for cell surface EGFR (A) or intracellular EGFR (B) with cetuximab, AF1280, 7A7 and 7A7 Fc Silent antibodies (blue peaks) or isotype control antibodies (red peaks) as described in Materials and Methods.
He, X. , Cruz, J. L. , Joseph, S. , Pett, N. , Chew, H. Y. , & Tuong, Z. K. , et al. (2018). Characterization of 7a7, an anti-mouse egfr monoclonal antibody proposed to be the mouse equivalent of cetuximab. Oncotarget, 9(15), 12250.
Figure 3 7A7 and 7A7 Fc Silent do not detect EGFR expression by Western blot.
Protein was extracted from A431, MCF-7, 3T3-L1, NIH-3T3, HPV38, and TC-1 cells, and immunoblotted for EGFR (A–B) and β-tubulin (C–D). (A) AF1280; (B) 7A7 and 7A7 Fc Silent; (C) and (D) β-tubulin internal loading controls for (A) and (B) respectively. EGFR was detected as a ~ 170kDa band by AF1280 (as shown by arrow). Numbers at the bottom of each lane in (A) and (B) represent quantification of each lane normalised to corresponding β-tubulin measurements and relative to the highest resulting ratio on the respective blots. * = position of non-specific band used as a comparator for EGFR-specificity. E/β-tub; EGFR/β-tubulin, N/β-tub; non-specific band/β-tubulin.
He, X. , Cruz, J. L. , Joseph, S. , Pett, N. , Chew, H. Y. , & Tuong, Z. K. , et al. (2018). Characterization of 7a7, an anti-mouse egfr monoclonal antibody proposed to be the mouse equivalent of cetuximab. Oncotarget, 9(15), 12250.
Figure 4 7A7 and 7A7 Fc Silent do not detect EGFR expression by immunofluorescence.
Four cell lines, A431, 3T3-L1, HPV38 and TC-1 were used to assess the binding of cetuximab (A), AF1280 (B), 7A7 (C) and 7A7 Fc Silent (D) antibodies as described in Materials and Methods. Red staining: Alexa Fluor 594-conjugated secondary antibodies, Blue staining: DAPI (nucleus), Green staining: plasma membrane as indicated by wheat germ agglutinin (WGA-FITC). Cmab; Cetuximab. Scale bar is 20 μm.
He, X. , Cruz, J. L. , Joseph, S. , Pett, N. , Chew, H. Y. , & Tuong, Z. K. , et al. (2018). Characterization of 7a7, an anti-mouse egfr monoclonal antibody proposed to be the mouse equivalent of cetuximab. Oncotarget, 9(15), 12250.
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PNBL-017 | Recombinant Anti-Human EGFR VHH Single Domain Antibody (PNBL-017) | FuncS, ELISA, IF | Llama VHH |
PNBL-018 | Recombinant Anti-Human EGFR VHH Single Domain Antibody (PNBL-018) | FuncS, SPR | Llama VHH |
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(Creative Biolabs Cat# PABW-088, RRID: AB_3111682)
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