Mouse anti-Human EGFR Antibody | anti-EGFR antibody

Anti-EGFR [Matuzumab], Mouse IgG1

Cat. Num. AAA72239

• Gene Names: EGFR; ERBB; HER1; mENA; ERBB1; PIG61
• Reactivity: Human
• Applications: Western Blot, Immunofluorescence, Flow Cytometry, Functional Assay, ELISA
• Purity: Protein A affinity purified
• Synonyms: EGFR; N/A; Anti-EGFR [Matuzumab], Mouse IgG1; Anti-EGFR [Matuzumab]; ErbB-1; ERBB1; HER1; EGF; Epidermal growth factor receptor; Proto-oncogene c-ErbB-1; Receptor tyrosine-protein kinase erbB-1; EMD72000; mAb425; 425; mAb 425; anti-EGFR antibody

• Host: Mouse
• Reactivity: Human
• Isotype: Mouse IgG1, Kappa
• Clone Number: Matuzumab
• Specificity: This antibody binds the extracellular domain III of the human EGFR and does not cross react with murine EGFR. It was also reported that residues S460/G461 in EGFR domain III are essential components of the epitope. This antibody binds at a site distinct from the EGF binding site, preventing conformational re-arrangement required for dimerization, a process essential for high-affinitiy ligand binding (Schmiedel, 2008). This antibody can only recognize the active form of EGFR. The antibody binds to both glyco- and aglyco receptor forms. EGFR is a transmembrane mitogenic glycoprotein with a cytoplasmically facing tyrosine kinase site which is activated by binding of the epidermal growth factor in the external domain. It also binds transforming growth factor alpha (TGFa). EGFR a member of the epidermal growth factor family and is closely related to receptor tyrosine kinases. This receptor tyrosine kinase binds ligands of the EGF family and activates signallying cascades to convert extracellular cues into appropriate cellular responses. Mutations that lead to EGFR overexpression (known as upregulation) or overactivity have been associated with a number of cancers, including lung cancer, anal cancers and glioblastoma multiforme as well as inflammatory diseases.
• Purity/Purification: Protein A affinity purified
• Form/Format: PBS with 0.02% Proclin 300.
• Concentration: 1mg/ml (varies by lot)

• Applicable Applications for anti-EGFR antibody: WB (Western Blot), IF (Immunofluorescence), FCM/FACS (Flow Cytometry), ELISA, Blocking, Antagonistic Antibody
• Immunogen: The parental mouse antibody was generated by immunizing BALB/c mice intraperitoneally with A431 cells in phosphate buffered saline (PBS). Later on the humanzied version of the antibody was created by grafting CDRs of the murine antibody onto human constant regions.
• Chimeric Use Statement: This chimeric murine antibody was made using the variable domain sequences of the original Human IgG1 format, for improved compatibility with existing reagents, assays and techniques.
• Preparation and Storage: Store at 4 degree C for up to 3 months. For longer storage, aliquot and store at -20 degree C.

ELISA

(Binding curve of anti-EGFR antibody Matuzumab (AAA72239-10.0) to EGFR-Fc fusion protein ELISA Plate coated with human EGFR-Fc (Pr00117-10.9) at a concentration of 5 ug/ml. A 3-fold serial dilution from 100 to 0.1 ng/ml was performed using AAA72239-10.0. For detection, a 1:4000 dilution of HRP labelled anti-human kappa light chain antibody was used.)

IF (Immunofluorescence)

(Immunofluorescence staining of fixed A431 cells with anti-EGFR antibody Matuzumab (AAA72239) Immunofluorescence analysis of unpermeabilisd paraformaldehyde fixed A431 cells on Shi-fix coverslips stained with the chimeric rabbit version of Matuzumab (AAA72239-23.0) at 10 ug/ml for 1h followed by Alexa Fluor 488 secondary antibody (1 ug/ml), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom AAA72239-23.0, DAPI, merged channels and an isotype control. The isotype control was stained with unimmunised rabbit IgG followed by Alexa Fluor 488 secondary antibody.)

FCM/FACS (Flow Cytometry)

(Flow-cytometry using the anti-EGFR antibody Matuzumab (AAA72239) HeLa (A) and A431 (B) cells were stained with unimmunized rabbit IgG antibody (black line) or the rabbit-chimeric version of Matuzumab (AAA72239-23.0, blue line) at a concentration of 10 ug/ml for 30 mins at RT. After washing, bound antibody was detected using an anti-rabbit IgG JK (FITC-conjugate) antibody (129936) at 2 ug/ml and cells analyzed on a FACSCanto flow-cytometer.)

IHC (Immunohiostchemistry)

(Immunohistochemical staining of rat stomach tissue using anti-EGFR antibody (AAA72239) Matuzumab. Anti-EGFR staining of formaldehyde fixed paraffin embedded rat stomach tissue, at 40x magnification. The human IgG1-chimeric version of Matuzumab (AAA72239-10.0) was used to stain samples at a concentration of 5 ug/ml.)

IHC (Immunohistochemistry)

(Immunohistochemical staining of mouse skin tissue using anti-EGFR antibody (AAA72239) Matuzumab. Anti-EGFR staining of formaldehyde fixed paraffin embedded mouse skin tissue, at 40x magnification. The human IgG1-chimeric version of Matuzumab (AAA72239-10.0) was used to stain samples at a concentration of 5 ug/ml.)

Related Product Information for anti-EGFR antibody

This antibody is derived from a mouse parental clone mAb 425 which has demonstrated anti-tumor activity against solid tumors in phase I clinical trials. The binding characterization of this antibody to human EGFR was done using ELISA. This antibody was also found to bind human EGFR with a similar avidity as the parental mouse antibody 425 (PMID: 1798701). The parental mouse antibody 425 in combination with C225 (Cetuximab) reduced growth and survival of EGFR overexpressing MDA-MB-468 breast cancer cells more effectively than either antibody alone. The combination was also reported to effectively inhibited AKT and MAPK phosphorylation in MDA-MB-468 cell (PMID: 18424917). This antibody was used in a phase 1 study to investigate the safety and tolerability and to explore the pharmacokinetic and pharmacodynamic profile in patients with solid tumors that express EGFR (PMID: 14701780). This antibody has been used in various phase I studies alone or in combination with other therapeutic agents to treat patients with various type of cancers (PMID: 16533873; 16622465; 19238629). A study reported that the antitumor effects of matuzumab and cetuximab depend on inhibition of EGFR downstream signaling mediated by Akt or Erk rather than on inhibition of EGFR itself (PMID: 18033688).

Product Categories/Family for anti-EGFR antibody

tumor marker; biosimilar; immunotherapy; cancer; oncology; tumor; glioma

References

Kettleborough et al. Humanization of a mouse monoclonal antibody by CDR-grafting: the importance of framework residues on loop conformation. Protein Eng. 1991 Oct;4(7):773-83. PMID:1798701

NCBI and Uniprot Product Information

• NCBI GI #: 29725609
• NCBI GeneID: 1956
• NCBI Accession #: NP_005219.2
• NCBI GenBank Nucleotide #: NM_005228.3
• UniProt Accession #: P00533; O00688; O00732; P06268; Q14225; Q68GS5; Q92795; Q9BZS2; Q9GZX1; Q9H2C9; Q9H3C9; Q9UMD7
• Molecular Weight: 134,277 Da
• NCBI Official Full Name: epidermal growth factor receptor isoform a
• NCBI Official Synonym Full Names: epidermal growth factor receptor
• NCBI Official Symbol: EGFR
• NCBI Official Synonym Symbols: ERBB; HER1; mENA; ERBB1; PIG61
• NCBI Protein Information: epidermal growth factor receptor; proto-oncogene c-ErbB-1; cell growth inhibiting protein 40; cell proliferation-inducing protein 61; receptor tyrosine-protein kinase erbB-1; avian erythroblastic leukemia viral (v-erb-b) oncogene homolog
• UniProt Protein Name: Epidermal growth factor receptor
• UniProt Gene Name: EGFR
• UniProt Synonym Gene Names: ERBB; ERBB1; HER1
• UniProt Entry Name: EGFR_HUMAN