Human Anti-EGFR Recombinant Antibody (clone Cetuximab (IMC-C225)) (CAT#: PABW-039)

Recombinant Chimeric (Human/Mouse) Antibody (Cetuximab (IMC-C225) is capable of binding to EGFR, expressed in Chinese Hamster Ovary cells (CHO). This antibody was used in treating advanced-stage EGFR-expressing colorectal cancer.

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Figure 1 Immunoblot of EGFR and tyrosine-phosphorylated EGFR from pancreatic carcinoma cells, BxPC-3, and MiaPaCa-2.

The EGFR-overexpressing human epidermoid cell line A431 was used as a positive control. Total cellular lysates were collected from cells treated for 24 h with IMC-C225, then exposed to EGF for 5 min. Immunoblot analysis for EGFR expression showed a low but detectable level of receptor associated with the pancreatic cell lines. IMC-C225 treatment did not have any effect on EGFR protein levels; however, the antibody did block ligand binding as detected by the dramatic reduction in EGF-induced tyrosine phosphorylation of EGFR. Actin protein was used as a control for protein loading.

Buchsbaum, D. J., Bonner, J. A., Grizzle, W. E., Stackhouse, M. A., Carpenter, M., Hicklin, D. J., ... & Raisch, K. P. (2002). Treatment of pancreatic cancer xenografts with Erbitux (IMC-C225) anti-EGFR antibody, gemcitabine, and radiation. International Journal of Radiation Oncology* Biology* Physics, 54(4), 1180-1193.

Figure 2 In vitro inhibition of pancreatic carcinoma cell growth.

BxPC-3 and MiaPaCa-2 cells were treated with IMC-C225 (C225) on Day 0, gemcitabine (Gem) on Day 1 for 24 h, followed by 60Co irradiation (RT) on Day 2, or the combination of each agent, then counted on Day 4. BxPC-3 cells treated with the combination of all three agents were significantly inhibited in cell proliferation as compared to the untreated control cells (p<0.05). Similar results were observed with the MiaPaCa-2 cells, with the three-agent combination showing a significant inhibition of cell proliferation as compared to the untreated and IMC-C225 treatment groups. Data points are the average SEM of three independent experiments, each done in quadruplicate cultures, then normalized to the untreated control (100%).

Buchsbaum, D. J., Bonner, J. A., Grizzle, W. E., Stackhouse, M. A., Carpenter, M., Hicklin, D. J., ... & Raisch, K. P. (2002). Treatment of pancreatic cancer xenografts with Erbitux (IMC-C225) anti-EGFR antibody, gemcitabine, and radiation. International Journal of Radiation Oncology* Biology* Physics, 54(4), 1180-1193.

Figure 3 Analysis of pancreatic carcinoma cells for the induction of early apoptosis. Floating and adherent cells were collected 4 days posttreatment, stained with annexin V-FITC and propidium iodide, then analyzed by FACS using CellQuest software.

A significant increase in apoptosis was observed with the BxPC-3 cells after treatment with the three-agent combination as compared to the untreated, radiation (RT), IMC-C225, gemcitabine (Gem), and radiation C225 treatment groups (p<0.001). The BxPC-3 cells showed a significant increase in apoptotic cell death after treatment with Gem C225 and Gem radiation as compared to the untreated and radiation treatment groups (p <0.001). With MiaPaCa-2 cells, the three-agent combination and the two-agent combination of Gem radiation showed a significant increase in apoptotic cells as compared to all other treatment groups (p<0.001). Also the MiaPaCa-2 cells treated with Gem or Gem C225 showed a significant increase in apoptosis as compared to untreated, C225, radiation, and C225 radiation treatment groups (p<0.001). Data points are the average SEM of three independent experiments done in triplicate (n=9).

Buchsbaum, D. J., Bonner, J. A., Grizzle, W. E., Stackhouse, M. A., Carpenter, M., Hicklin, D. J., ... & Raisch, K. P. (2002). Treatment of pancreatic cancer xenografts with Erbitux (IMC-C225) anti-EGFR antibody, gemcitabine, and radiation. International Journal of Radiation Oncology* Biology* Physics, 54(4), 1180-1193.

Figure 4 TUNEL result of IMC-C225.

(A) An apparently viable area of a MiaPaCa-2 tumor xenograft from an untreated nude mouse; about 5%–10% of cells demonstrate apoptosis identified by the TUNEL technique. (B) A viable area of a MiaPaCa-2 xenograft tumor from a mouse treated for 1 week with IMC-C225 (1 mg every 3 days 2), gemcitabine (120 mg/kg 1 day after the first dose of mAb), and radiation (3 Gy at 1 day after gemcitabine); at 4 days after radiation, the mice were killed. About 30%–40% of cells exhibit apoptosis that is higher compared to the basal level in untreated tumors. (C) Untreated mouse with a MiaPaCa-2 tumor xenograft was injected i.p. with BrdU and killed 2 h later; about 70%–80% of the cells were
undergoing proliferation. (D) A marked reduction in proliferation as detected by BrdU staining in treated MiaPaCa-2 tumors (10%–15%) compared to the basal level in untreated tumors (60%–70%).

Buchsbaum, D. J., Bonner, J. A., Grizzle, W. E., Stackhouse, M. A., Carpenter, M., Hicklin, D. J., ... & Raisch, K. P. (2002). Treatment of pancreatic cancer xenografts with Erbitux (IMC-C225) anti-EGFR antibody, gemcitabine, and radiation. International Journal of Radiation Oncology* Biology* Physics, 54(4), 1180-1193.


Specifications

  • Immunogen
  • Human epidermal growth factor receptor
  • Host Species
  • Human
  • Derivation
  • Chimeric (mouse/human)
  • Type
  • Chimeric (mouse/human) IgG
  • Specificity
  • Human EGFR
  • Species Reactivity
  • Human
  • Clone
  • Cetuximab (IMC-C225)
  • Applications
  • FuncS

Applications

  • Application Notes
  • The EGFR antibody has been reported in applications of Western Blot, Inhibition, Flow Cytometry, TUNEL.

Target

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

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.

See other products for "EGFR"

* Abbreviations
3D IHC3D Immunohistochemistry
ActivActivation
AgonistAgonist
ApopApoptosis
BABioassay
BIBioimaging
BlockBlocking
Cell ScreeningCell Screening
SeparationCell Separation
ChIPChromatin Immunoprecipitation
CMCDComplement Mediated Cell Depletion
CostimCostimulation
CytCytotoxicity
DepletionDepletion
DBDot Blot
EMElectron Microscopy
ELISAEnzyme-linked Immunosorbent Assay
ELISPOTEnzyme-linked Immunosorbent Spot
FCFlow Cytometry
FuncSFunctional Assay
GSGel Super Shift Assay
HAHemagglutination
IAImmunoassay
IBImmunoblotting
ICCImmunocytochemistry
IDImmunodiffusion
IFImmunofluorescence
IHCImmunohistochemistry
IHC-FrImmunohistochemistry-Frozen
IHC-PImmunohistochemistry-Paraffin
REImmunohistology - Resin Sections
IPImmunoprecipitation
IRMAImmunoradiometric Assay
SHIn situ hybridization
InhibInhibition
ICFCIntracellular Staining for Flow Cytometry
KO/KD-WBKnockout/Knockdown target confirmation by Western Blot
Live cell imagingLive cell imaging
CyTOF®Mass Cytometry
MeDIPMethylated DNA Immunoprecipitation
MultiplexMultiplex bead-based assay
NeutNeutralization
PPProtein Purification
PGProteogenomics
RIRadial Immunodiffusion
RIARadioimmunoassay
StimStimulation
SPRSurface Plasmon Resonance
TCTissue Culture
TBTurbidimetry
WBWestern Blot

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