Anti-Human EGFR Therapeutic Antibody (cetuximab) (CAT#: TAB-0225CL)

Cetuximab is a recombinant chimeric monoclonal antibody that binds to the extracelluar domian of the human epidermal growth factor receptor, and it was used for the treatment of metastatic colorectal cancer, metastatic non-small cell lung cancer and head and neck cancer. The antibody derived from Cetuximab without the glycosylation of galactose-alpha-l,3-galactose which has ben shown to cause allergic responses in some patient.

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FuncS

Figure 1 Effects of cetuximab (C225) on tumor cell growth.

Du145, PC-3 and A431 cells were incubated with 100 μg/ml cetuximab for up to 72h. Control cells received no treatment. Cell number was counted using trypan blue viability assay and normalized to the pretreatment value.

Dhupkar, P., Dowling, M., Cengel, K., & Chen, B. (2010). Effects of anti-EGFR antibody cetuximab on androgen-independent prostate cancer cells. Anticancer research, 30(6), 1905-1910.

FuncS

Figure 2 Representative flow cytometry histograms showing the population of cells in M1 (Sub-G1), M2 (G0/G1), M3 (S) and M4 (G2-M) phases.

Du145, PC-3 and A431 cells were incubated with 100 μg/ml cetuximab (C225) for 72 h. Control cells received no treatment. At the end of treatment, cells were collected and prepared. Cellular DNA content was analyzed with flow cytometry after staining.

Dhupkar, P., Dowling, M., Cengel, K., & Chen, B. (2010). Effects of anti-EGFR antibody cetuximab on androgen-independent prostate cancer cells. Anticancer research, 30(6), 1905-1910.

FuncS

Figure 3 Western blots showing the effects of certuximab (C225) on EGFR signaling.

Du145, PC-3 and A431 cells were incubated with 100 μg/ml cetuximab (C225) for 72 h. Control cells received no treatment. At the end of treatment, cells were lysed and prepared. Cell samples were resolved by SDS-PAGE.

Dhupkar, P., Dowling, M., Cengel, K., & Chen, B. (2010). Effects of anti-EGFR antibody cetuximab on androgen-independent prostate cancer cells. Anticancer research, 30(6), 1905-1910.

Inhib

Figure 4 The cetuximab Fab fragment binds to sEGFR and inhibits sEGFR binding to EGF

A: Surface Plasmon Resonance (SPR) analysis of sEGFR binding to immobilized FabC225. A series of sEGFR samples of the indicated concentrations or of the domain III fragment of sEGFR (sEGFR-III) were passed over a Biosensor surface to which FabC225 had been covalently coupled. A representative data set of the equilibrium SPR response for each sample, expressed as the fraction of the maximum binding, is plotted as a function of the concentration of sEGFR. The inset shows that no additional binding is seen at higher concentrations. These data can be fit to a simple one-site Langmuir binding equation. The curve indicates the fit to the particular data set shown. KD values are the mean of at least three independent binding experiments. KD values are 2.3 ± 0.5 and 1.7 ± 0.6 nM for sEGFR and sEGFR-III, respectively. B: SPR equilibrium binding analysis, as described in A, of a similar set of samples of sEGFR and sEGFR-III to immobilized EGF. KD values are 130 ± 3 nM and 2.3 ± 0.5 μM for sEGFR and sEGFR-III, respectively. C: The ability of FabC225 to compete with immobilized EGF for sEGFR binding is shown. The indicated molar excesses of Fab were added to samples of a fixed concentration of sEGFR (600 nM). The equilibrium SPR responses obtained by passing these mixtures over immobilized EGF, expressed as a fraction of the response with no added Fab, are plotted as a function of the molar excess of Fab. Each data point is the mean of at least three independent measurements. All binding is abolished at a 1:1 stochiometry of FabC225:sEGFR, and the IC50 value for these conditions is 500 nM.

Li, S., Schmitz, K. R., Jeffrey, P. D., Wiltzius, J. J., Kussie, P., & Ferguson, K. M. (2005). Structural basis for inhibition of the epidermal growth factor receptor by cetuximab. Cancer cell, 7(4), 301-311.

Figure 5 shows the toxicity of cetuximab mutant antibody, 6-LC, (having reduced affinity to make it a partially antagonistic against EGFR) is potentiated by conjugation to payload via non-cleavable linker (6LC-DM1).

Figure 6 shows that partially antagonistic anti-EGFR antibody by IMGN (J2989A) is potentiated by conjugation (IMGN289) on normal keratinocytes.


Specifications

  • Host Species
  • Human
  • Type
  • Chimeric (mouse/human)
  • Specificity
  • Human
  • Clone
  • cetuximab
  • Applications
  • Inhib, FuncS
  • Related Disease
  • Metastatic colorectal cancer, metastatic non-small cell lung cancer, head and neck cancer

Applications

  • Application Notes
  • The antibody was validated for Inhibition, Function Assay. For details, refer to Published Data.

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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