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

Recombinant Mouse Antibody (7A7) is capable of binding to Mouse 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.

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.

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.

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.

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

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
  • EGFR; epidermal growth factor receptor; Wa5; wa2; Erbb; Errp; wa-2; Errb1; AI552599; 9030024J15Rik; avian erythroblastic leukemia viral (v-erb-b) oncogene homolog; waved 2

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"

See other products for "Clone 7A7"


For Research Use Only. Not For Clinical Use.

* 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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