Mouse Anti-EGFR Recombinant Antibody (clone 7A7) (CAT#: PABW-088)

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.

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

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.

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.

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.

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.


Specifications

  • Immunogen
  • Human epidermal growth factor receptor
  • Host Species
  • Mouse
  • Type
  • Mouse IgG
  • Specificity
  • Human EGFR
  • Species Reactivity
  • Human
  • Clone
  • 7A7
  • Applications
  • ELISA, WB

Product Property

  • Purity
  • >95% as determined by analysis by SDS-PAGE
  • Concentration
  • Please refer to the vial label for the specific concentration.
  • Storage
  • Centrifuge briefly prior to opening vial. Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freeze/thaw cycles.

Applications

  • Application Notes
  • The EGFR antibody has been reported in applications of Enzyme-linked Immunosorbent Assay, Flow Cytometry, Western Blot, Immunofluorescence. Flow Cytometry: Cells were prepared for flow cytometry by washing with PBS and then FACs buffer (2% FBS in PBS). Cell suspensions (1x10^6cells) were stained with 7A7, 7A7 Fc Silent, cetuximab or AF1280 antibodies, and incubated with Alexa 488-conjugated-anti-mouse, anti-human or anti-goat secondary antibodies respectively. Immunofluorescence: Sections were then incubated with either 7A7, 7A7 Fc Silent, cetuximab or AF1280 antibody overnight at 4°C. After washing with PBS, sections were incubated with Alexa-Fluor-594 conjugated anti-mouse, anti-human or anti-goat secondary antibodies respectively. Western Blot: PVDF membranes were blocked with 2% BSA in PBS and incubated with 7A7, 7A7 Fc Silent, anti-mouse EGFR "AF1280" or β-tubulin primary antibodies and anti-mouse or anti-goat HRP secondary antibodies.

Target

  • Alternative Names
  • ERBB; ERRP; HER1; mENA; ERBB1; PIG61; NISBD2; epidermal growth factor receptor; EGFR vIII; avian erythroblastic leukemia viral (v-erb-b) oncogene homolog; cell growth inhibiting protein 40; cell proliferation-inducing protein 61; epidermal growth factor receptor tyrosine kinase domain; erb-b2 receptor tyrosine kinase 1; proto-oncogene c-ErbB-1; receptor tyrosine-protein kinase erbB-1

Related Resources

  • Related Diseases
  • Related Signaling Pathways

Product Notes

This is a product of Creative Biolabs' Hi-Affi™ recombinant antibody portfolio, which has several benefits including:

• Increased sensitivity
• Confirmed specificity
• High repeatability
• Excellent batch-to-batch consistency
• Sustainable supply
• Animal-free production

See more details about Hi-Affi™ recombinant antibody benefits.

Downloads

Download resources about recombinant antibody development and antibody engineering to boost your research.

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Fc Glycosylation

High-mannose Glycoform

CAT Product Name Application Type
Gly-055LC-1 Recombinant Anti-Human EGFR Antibody (Fc glycosylation/High-mannose glycosylated) ELISA Chimeric antibody (mouse/human)

Deglycosylated Antibody (Non-glycosylated IgGs)

CAT Product Name Application Type
Gly-167LC Recombinant Anti-Human EGFR Antibody (Non-glycosylated) ELISA Human antibody

Chicken IgY Antibody

CAT Product Name Application Type
BRD-0183MZ Chicken Anti-EGFR Polyclonal IgY WB Chicken antibody
BRD-0668MZ Chicken Anti-EGFR Polyclonal IgY WB Chicken antibody

Neutralizing Antibody

ADCC Enhanced Antibody

CAT Product Name Application Type
AFC-TAB-165 Afuco™ Anti-EGFR ADCC Recombinant Antibody (Matuzumab), ADCC Enhanced Neut, ELISA, IF, IP, FuncS, FC ADCC enhanced antibody
AFC-TAB-464CQ Afuco™ Anti-EGFR ADCC Recombinant Antibody (Tomuzotuximab), ADCC Enhanced ELISA, IHC, FC, IP, IF, FuncS ADCC enhanced antibody
AFC-TAB-003 Afuco™ Anti-EGFR ADCC Recombinant Antibody (Cetuximab), ADCC Enhanced IF, IP, Neut, FuncS, ELISA, FC ADCC enhanced antibody
AFC-TAB-040 Afuco™ Anti-EGFR ADCC Recombinant Antibody (Zalutumumab), ADCC Enhanced ELISA, FC, IP, FuncS, IF, Neut ADCC enhanced antibody
AFC-TAB-119 Afuco™ Anti-EGFR ADCC Recombinant Antibody (Necitumumab), ADCC Enhanced FC, IP, ELISA, Neut, FuncS, IF ADCC enhanced antibody

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For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.

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