Cetuximab (anti-EGFR)

Catalog No.A2000 Batch: A200007

Technical Data

CAS No.	205923-56-4
Formulation	100 mM Pro-Ac, 20 mM Arg, pH5.0
Isotype	Human IgG1
Source	CHO cells
Storage (From the date of receipt)	Store the undiluted solution at 4°C in the dark to avoid freeze-thaw cycles
Purity	99.81%
Protein concentration	3.63 mg/ml
Endotoxin Level	<1 EU/mg

Biological Activity

Description

Cetuximab, a novel molecular-targeted agent,is an inhibitor of EGFR monoclonal humanized antibody interacting with the extracellular binding site of EGFR to block ligand stimulation. MW：145.781 KD.

Targets

EGFR ^[4]
(Cell-free assay)

0.39 nM(Kd)

In vitro

Cetuximab treatment increases mitochondrial priming of EGFR-expressing HeLa cells but not in EGFR-expressing MDA-MD-231 cells.^[2]

In vivo

C225 enhanced the antitumor activity of several chemotherapeutic drugs in mouse xenograft models^[1]. Cetuximab, exerts its antitumor efficacy by multiple mechanisms that include the inhibition of cell cycle progression by arrest in the G1- phase and decreased cell number in the S-phase. Cell cycle arrest in the G1-phase also induces apoptosis by the induction and activation of proapoptotic molecules. cetuximab alone and in synergy with carboplatin resulted in decreases of tumor size, metastatic spread, and MVD in NCI-N87 tumors with EGFR cell surface expression and absence of mutations in BRAF and K-ras, whereas cetuximab had minimal in vitro effect and no in vivo treatment efficacy in tumors derived from MKN-45, in which the phenotype was also BRAF and K-ras wildtype, but which had only weak cytoplasmic EGFR protein expression^[3].

Protocol (Only for Reference)

Cell Assay:	Cells were lysed at a density of 1 x 10⁶/50 μL in lysis buffer (0.25 M Tris-HCl, 2% sodium dodecylsulfate, 4% β-mercaptoethanol, 10% glycerol, 0.02% bromophenol blue) supplemented with 1 X protease/phosphatase inhibitor cocktail. Cell lysates were then loaded onto polyacrylamide gels with sodium dodecyl sulfate. After electrophoresis, proteins were transferred to polyvinylidene difluoride (PVDF) membranes. The transblotted membranes were blocked for 1 hr and then probed with appropriate primary antibodies overnight at 4 ℃. Next day, the membranes were washed three times for a total of 30 min and then incubated with IRDye 680RD Donkey anti-Rabbit IgG (H + L) or IRDye 800CW Donkey anti-Mouse IgG (H + L) in darkness at room temperature for 1 h. After another three washes, scan immunoblot membranes and quantify band intensity.
Animal Study:	Objective: Antitumor activity of cetuximab in murine gastric cancer model Animal Models: Nude mouse model for human gastric cancer (CD-1/nu-nu mice) Formulation: 0.9% NaCl Dosages: 1 mg/kg Administration: i.p. Reference: https://www.ncbi.nlm.nih.gov/pubmed/22011788 Objective: To investigate the relationship between the EGFR levels and the responsiveness to cetuximab treatment in human cancer xenograft models Animal Models: Female athymic, nude mice were implanted s.c. with ~1 mm3 tumor fragments Formulation: PBS Dosages: 0.25, 0.5 or 1 mg/mouse Administration: i.p. Reference: https://www.ncbi.nlm.nih.gov/pubmed/27186886 Cetuximab can apply to nude mice, various cancer cell lines and other related assays (Only for Reference)

Cell Assay:

Cells were lysed at a density of 1 x 10⁶/50 μL in lysis buffer (0.25 M Tris-HCl, 2% sodium dodecylsulfate, 4% β-mercaptoethanol, 10% glycerol, 0.02% bromophenol blue) supplemented with 1 X protease/phosphatase inhibitor cocktail. Cell lysates were then loaded onto polyacrylamide gels with sodium dodecyl sulfate. After electrophoresis, proteins were transferred to polyvinylidene difluoride (PVDF) membranes. The transblotted membranes were blocked for 1 hr and then probed with appropriate primary antibodies overnight at 4 ℃. Next day, the membranes were washed three times for a total of 30 min and then incubated with IRDye 680RD Donkey anti-Rabbit IgG (H + L) or IRDye 800CW Donkey anti-Mouse IgG (H + L) in darkness at room temperature for 1 h. After another three washes, scan immunoblot membranes and quantify band intensity.

Animal Study:

Objective: Antitumor activity of cetuximab in murine gastric cancer model
Animal Models: Nude mouse model for human gastric cancer (CD-1/nu-nu mice)
Formulation: 0.9% NaCl
Dosages: 1 mg/kg
Administration: i.p.
Reference: https://www.ncbi.nlm.nih.gov/pubmed/22011788

Objective: To investigate the relationship between the EGFR levels and the responsiveness to cetuximab treatment in human cancer xenograft models
Animal Models: Female athymic, nude mice were implanted s.c. with ~1 mm3 tumor fragments
Formulation: PBS
Dosages: 0.25, 0.5 or 1 mg/mouse
Administration: i.p.
Reference: https://www.ncbi.nlm.nih.gov/pubmed/27186886

Cetuximab can apply to nude mice, various cancer cell lines and other related assays (Only for Reference)

References	[1] Gu J, et al. Biochem Res Int. 2016, 2016:7016907. [2] Pan R, et al. Cell. 2022 Apr 28;185(9):1521-1538.e18. [3] Hotz B, et al. Gastric Cancer. 2012, 15(3):252-64.

References

Customer Product Validation

Data from [Data independently produced by , , J Thorac Oncol, 2018, 13(6):810-820]

Mice showed progression disease to first-line treatment were randomized 1:1 to the two arms of treatment and were treated until onset of progression disease or until the end-time of experiment in case of response (fixed to 52 weeks from the start of first-line therapy ), as indicated in in the Materials and Methods section. Growth curves of HCC827 xenografts treated in secondline with osimertinib plus selumetinib or cetuximab are represented as changes in the values of volumes as percentage compared to baseline tumor volume at the time of progression diasease to first-line (defined as 100%) for each case. Median tumor volume at PD2 was of 348 mm³.

Data from [Data independently produced by , , ]

Selleck's Cetuximab (anti-EGFR) has been cited by 50 publications

Base editing screens define the genetic landscape of cancer drug resistance mechanisms [ Nat Genet, 2024, 10.1038/s41588-024-01948-8]	PubMed: 39424923
The effector program of human CD8 T cells supports tissue remodeling [ J Exp Med, 2024, 221(2)e20230488]	PubMed: 38226976
EGFR and HER2 hyper-activation mediates resistance to endocrine therapy and CDK4/6 inhibitors in ER+ breast cancer [ Cancer Lett, 2024, 593:216968]	PubMed: 38788968
Label-free mapping of cetuximab in multi-layered tumor oral mucosa models by atomic force-microscopy-based infrared spectroscopy [ Analyst, 2024, 149(7):2122-2130]	PubMed: 38436119
Label-free mapping of cetuximab in multi-layered tumor oral mucosa models by atomic force-microscopy-based infrared spectroscopy [ Analyst, 2024, 149(7):2122-2130]	PubMed: 38436119
CDK4/6 inhibition to resensitize BRAF/EGFR inhibitor in patient-derived BRAF/PTEN-mutant colon cancer cells [ Transl Cancer Res, 2024, 13(7):3695-3703]	PubMed: 39145064
Differential Regulation of the STING Pathway in Human Papillomavirus-Positive and -Negative Head and Neck Cancers [ Cancer Res Commun, 2024, 4(1):118-133]	PubMed: 38147007
Double Digital Assay for Single Extracellular Vesicle and Single Molecule Detection [ Adv Sci (Weinh), 2023, 10(33):e2303619]	PubMed: 37802976
Feedback activation of EGFR/wild-type RAS signaling axis limits KRASG12D inhibitor efficacy in KRASG12D-mutated colorectal cancer [ Oncogene, 2023, 42(20):1620-1633]	PubMed: 37020035
EGFR-T790M Mutation-Derived Interactome Rerouted EGFR Translocation Contributing to Gefitinib Resistance in Non-Small Cell Lung Cancer [ Mol Cell Proteomics, 2023, 22(9):100624]	PubMed: 37495186

FOR RESEARCH USE ONLY. NOT FOR USE IN HUMANS.

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