Mouse Anti-EGFR Recombinant Antibody (clone 7A7)

Figure 1 7A7 binding to the EGFR. (A) Competition assay of 7A7 and Nimotuzumab.

ELISA plates were coated with 10 ug/ml eEGFR and incubated with mixtures containing a constant concentration of 5 ug/ml 7A7 and increasing concentrations of Nimotuzumab. Bound antibodies were developed by a secondary phosphatase conjugated anti-mouse or anti-human antibody, respectively. (B) No recognition of EGFR-D3 by 7A7. ELISA plates were coated with 10 ug/ml of EGFR-D3, and incubated with 7A7 or Nimotuzumab at 10 ug/ml.

Talavera, A. , Mackenzie, J. , Garrido, G. , Friemann, R. , Alejandro López-Requena, & Moreno, E. , et al. (2011). Structure of the fab fragment of the anti-murine egfr antibody 7a7 and exploration of its receptor binding site. Molecular Immunology, 48(12-13), 1578-1585.

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.

Figure 1 Anti-Human EGFR Antibody (PABW-088) in ELISA

ELISA analysis of PABW-088 was performed with EGFR Protein, Human, Recombinant (His Tag).
The secondary antibody: HRP-Anti-Mouse IgG (H+L)

Figure 2 Anti-Human EGFR Antibody (PABW-088) in DB

Dot blot analysis of PABW-088 was performed with EGFR Protein, Human, Recombinant (His Tag).
PABW-088 incubation concentration: 2ng/μL.
NC: Negative control PC: Positive control