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Rabbit Recombinant Monoclonal EGFR antibody. Suitable for WB, IP, Flow Cyt (Intra), IHC-P, ICC/IF and reacts with Transfected cell lysate - Human, Human, Transfected cell line - Human, Transfected cell line samples.

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFRvIII antibody [EPR28380-83] (AB313646), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFRvIII antibody [EPR28380-83] (AB313646), expandable thumbnail
  • Immunoprecipitation - Anti-EGFRvIII antibody [EPR28380-83] (AB313646), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-EGFRvIII antibody [EPR28380-83] (AB313646), expandable thumbnail
  • Western blot - Anti-EGFRvIII antibody [EPR28380-83] (AB313646), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBIPFlow Cyt (Intra)IHC-PICC/IF
Human
Expected
Expected
Expected
Tested
Expected
Transfected cell line
Not recommended
Not recommended
Not recommended
Tested
Not recommended
Transfected cell line - Human
Not recommended
Not recommended
Tested
Not recommended
Tested
Transfected cell lysate - Human
Tested
Tested
Not recommended
Not recommended
Not recommended

Tested
Tested

Species
Transfected cell lysate - Human
Dilution info
1/1000
Notes

-

Expected
Expected

Species
Human
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Transfected cell line - Human, Transfected cell line
Dilution info
-
Notes

-

Tested
Tested

Species
Transfected cell lysate - Human
Dilution info
1/30
Notes

-

Expected
Expected

Species
Human
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Transfected cell line - Human, Transfected cell line
Dilution info
-
Notes

-

Tested
Tested

Species
Transfected cell line - Human
Dilution info
1/500
Notes

-

Expected
Expected

Species
Human
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Transfected cell lysate - Human, Transfected cell line
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/100
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species
Transfected cell line
Dilution info
1/100
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species
Transfected cell lysate - Human, Transfected cell line - Human
Dilution info
-
Notes

-

Tested
Tested

Species
Transfected cell line - Human
Dilution info
1/500
Notes

-

Expected
Expected

Species
Human
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Transfected cell lysate - Human, Transfected cell line
Dilution info
-
Notes

-

Associated Products

Select an associated product type

1 product for Alternative Version

Target data

Function

The protein expressed by the EGFR gene functions as a receptor tyrosine kinase, binding with ligands from the EGF family to activate several signaling cascades that translate extracellular signals into cellular responses. Known ligands include EGF, TGFA/TGF-alpha, AREG, epigen, BTC/betacellulin, epiregulin, and HBEGF. Ligand binding triggers receptor dimerization and autophosphorylation, recruiting adapter proteins like GRB2 and activating complex downstream signaling pathways such as RAS-RAF-MEK-ERK, PI3 kinase-AKT, PLCgamma-PKC, and STATs, and possibly the NF-kappa-B cascade. EGFR also directly phosphorylates proteins like RGS16 and MUC1, influencing various cellular processes including its coupling with G protein-coupled receptor signaling and cell migration through interaction with CCDC88A/GIV. Isoform 2 of EGFR may act as an antagonist to EGF actions. In microbial infection, EGFR also serves as a receptor facilitating hepatitis C virus (HCV) entry into hepatocytes by promoting CD81-CLDN1 receptor complexes and enhancing membrane fusion with HCV envelope glycoproteins. This supplementary information is collated from multiple sources and compiled automatically.

Alternative names

ERBB, ERBB1, HER1, EGFR, Epidermal growth factor receptor, Proto-oncogene c-ErbB-1, Receptor tyrosine-protein kinase erbB-1

Recommended products

Rabbit Recombinant Monoclonal EGFR antibody. Suitable for WB, IP, Flow Cyt (Intra), IHC-P, ICC/IF and reacts with Transfected cell lysate - Human, Human, Transfected cell line - Human, Transfected cell line samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR28380-83
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

EGFR VIII also known as EGFRvIII EGFR of 83 or 83 EGFR is a mutant form of the epidermal growth factor receptor (EGFR) with a molecular weight of approximately 145 kDa. This variant results from a deletion of exons 2 to 7 in the EGFR gene which produces a truncated protein that lacks a portion of the extracellular ligand-binding domain. The EGFRvIII does not bind ligands and is constitutively active. It is often expressed in cancer cells particularly in glioblastomas and some other solid tumors but not in normal tissues.

Biological function summary

EGFRvIII modifies signaling pathways that promote cell survival and proliferation. It is a part of cell membrane-based complexes interacting with proteins like PI3K and Src which helps in activating downstream signaling cascades. This constitutive signaling leads to increased oncogenic potential allowing the cells to thrive in uncontrolled manner contributing to tumor progression and resistance to conventional therapies.

Pathways

EGFRvIII participates in key signaling pathways such as the PI3K/AKT pathway and the RAS/RAF/MEK/ERK pathway. These pathways are important in transmitting signals from receptor tyrosine kinases to the cell nucleus. EGFRvIII links with proteins like AKT and ERK to drive processes that control growth and survival. The dysregulation of these pathways due to EGFRvIII activity can result in enhanced proliferative and survival signals in cancer cells.

Associated diseases and disorders

EGFRvIII is strongly associated with glioblastoma and non-small cell lung cancer. In these conditions EGFRvIII expression correlates with poor prognosis and an aggressive tumor phenotype. The presence of EGFRvIII often accompanies other oncogenic proteins such as PTEN whose loss of function can amplify the oncogenic signaling of EGFRvIII. As a result targeting EGFRvIII represents a potential therapeutic intervention in such aggressive cancer types.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

9 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFRvIII antibody [EPR28380-83] (ab313646), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFRvIII antibody [EPR28380-83] (ab313646)

    EGFRvIII Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of Human glioblastoma tissue using rabbit Anti-EGFRvIII antibody

    Chromogenic duplex (2-plex) immunohistochemical staining of formalin fixed paraffin-embedded human glioblastoma tissue performed on a Roche Ventana Discovery Ultra instrument.

    Heat mediated antigen retrieval was performed with CC1 solution for 64 min at 95°C. Two rounds of primary antibody incubation were performed each for 16 min at 37°C. anti-EGFRvIII (ab313646; 1st primary antibody; 3 µg/ml) and anti-EGFR (Anti-EGFR antibody [EP38Y] ab52894; 2nd primary antibody; 0.5 µg/ml). Antibody denaturation was conducted after the first round of staining using Ultra CC2 solution for 8 min at 100°C to avoid cross reactivity.

    Signal was developed with anti-rabbit HQ followed by anti-HQ HRP coupled with purple (Discovery purple kit (RUO) #760-229) and teal (Discovery teal kit (RUO) #760-247) chromogens and counterstained with haematoxylin II.

    Tissue obtained from the Human Research Tissue Bank, supported by the NHR Cambridge Biomedical Research Centre.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFRvIII antibody [EPR28380-83] (ab313646), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFRvIII antibody [EPR28380-83] (ab313646)

    EGFRvIII Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of Human glioblastoma and placenta tissues using rabbit Anti-EGFRvIII antibody

    Immunohistochemical analysis of formalin fixed paraffin-embedded human glioblastoma and placenta tissues labelling EGFRvIII with ab313646 and EGFR with Anti-EGFR antibody [EP38Y] ab52894 at a concentration of 2.5 µg/ml and 0.5 µg/ml.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument with a Bond™ Polymer Refine Detection kit. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins. Primary antibodies were incubated for 30 mins at room temperature. Sections were counterstained with Hematoxylin.

    Tissue obtained from the Human Research Tissue Bank, supported by the NHR Cambridge Biomedical Research Centre.

  • Immunoprecipitation - Anti-EGFRvIII antibody [EPR28380-83] (ab313646), expandable thumbnail

    Immunoprecipitation - Anti-EGFRvIII antibody [EPR28380-83] (ab313646)

    EGFRvIII was immunoprecipitated from 0.35 mg of 293T (human embryonic kidney epithelial cell) transfected with a human EGFRvIII expression vector containing a His-tag using ab313646 at 1/30 dilution (2µg in 0.35mg whole cell lysate). Western blot was performed on the immunoprecipitate using ab313646 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution. Isotype control - Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730).

    Lanes 1-3: 293T (human embryonic kidney epithelial cell) transfected with a human EGFRvIII expression vector containing a His-tag

    Lane 1: whole cell lysate at 0.3µg

    Lane 2: IP with ab313646

    Lane 3: IP with Isotype control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730)

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    All lanes: Immunoprecipitation - Anti-EGFRvIII antibody [EPR28380-83] (ab313646) at 1/30 dilution

    All lanes: 293T (human embryonic kidney epithelial cell) transfected with a human EGFRvIII expression vector containing a His-tag whole cell lysate

    Secondary

    All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/1000 dilution

    Exposure time: 5s

  • Immunocytochemistry/ Immunofluorescence - Anti-EGFRvIII antibody [EPR28380-83] (ab313646), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-EGFRvIII antibody [EPR28380-83] (ab313646)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) labelling EGFRvIII with ab313646 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Cells were counterstained with anti-Myc tag Mouse mab [9E10] (Alexa Fluor® 594) at 1/100 dilution (Red). The Nuclear counterstain is DAPI (Blue).

    Confocal image showing cytoplasmic and membrane staining in 293T cells overexpressing human EGFRvIII. No staining in 293T cells overexpressing human wild-type EGFR. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/mL) dilution.

  • Western blot - Anti-EGFRvIII antibody [EPR28380-83] (ab313646), expandable thumbnail

    Western blot - Anti-EGFRvIII antibody [EPR28380-83] (ab313646)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST
    This antibody recognizes human EGFRvIII but not human wild-type EGFR. In western blot, anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution, anti-His antibody (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution, and anti-EGFR antibody staining at 1/1000 dilution.

    All lanes: Western blot - Anti-EGFRvIII antibody [EPR28380-83] (ab313646) at 1/1000 dilution

    Lane 1: 293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a His tag, whole cell lysate at 10 µg

    Lane 2: 293T transfected with a human EGFR expression vector containing a His tag, whole cell lysate at 10 µg

    Lane 3: 293T transfected with a human EGFRvIII expression vector containing a His tag, whole cell lysate at 10 µg

    Lane 4: 293T transfected with a human EGFRvIII expression vector containing a His tag, whole cell lysate at 2.5 µg

    Lane 5: 293T transfected with a human EGFRvIII expression vector containing a His tag, whole cell lysate at 0.62 µg

    Lane 6: 293T transfected with a human EGFRvIII expression vector containing a His tag, whole cell lysate at 0.31 µg

    Lane 7: 293T transfected with a human EGFRvIII expression vector containing a His tag, whole cell lysate at 0.16 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Observed band size: 134 kDa

    Exposure time: 5.5s

  • Flow Cytometry (Intracellular) - Anti-EGFRvIII antibody [EPR28380-83] (ab313646), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-EGFRvIII antibody [EPR28380-83] (ab313646)

    Flow cytometric analysis of 293T (human embryonic kidney epithelial cell) transfected with a human EGFRvIII expression vector containing a His-Myc tag (top left, bottom left); human wild-type EGFR expression vector containing a His-Myc tag (top right); empty vector containing a His-Myc tag (bottom right). Cells were fixed with 2% paraformaldehyde and permeabilised with 0.1% Tween-20. Cell labelling of EGFRvIII with ab313646 at 1/500 dilution. A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) was used at 1/5000 dilution. Isotype control - Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730). No cross-reactivity with human wild-type EGFR.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFRvIII antibody [EPR28380-83] (ab313646), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFRvIII antibody [EPR28380-83] (ab313646)

    Immunohistochemical analysis of paraffin-embedded 293T (human embryonic kidney epithelial cell) transfected with (A) human EGFRvIII expression vector containing a Myc-His tag; (B) human wild-type EGFR expression vector containing a Myc-His tag; (C) empty vector containing a His tag. Whole cell pellet labelling of EGFRvIII with ab313646 at 1/100 (5.09 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Positive staining on 293T overexpressing human EGFRvIII (image A). No staining on 293T overexpressing human wild-type EGFR (image B) or the empty vector control (image C). The section was incubated with ab313646 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFRvIII antibody [EPR28380-83] (ab313646), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFRvIII antibody [EPR28380-83] (ab313646)

    Immunohistochemical analysis of paraffin-embedded (A, D) human placenta; (B, E) human glioblastoma; (C, F) human lung adenocarcinoma tissue.

    A-C: Sections were incubated with anti-EGFR antibody [SP84] - C-terminal (Anti-EGFR antibody [SP84] - C-terminal ab227642; 1:100 ) for 30 mins at room temperature. Positive staining on human placenta (image A), glioblastoma (image B), and lung adenocarcinoma (image C). D-F: Sections were incubated with anti-EGFRvIII antibody [EPR28380-83] (ab313646; 1:100 [5.09 µg/mL]) for 30 mins at room temperature. Positive staining on human glioblastoma (image E), no staining on placenta (image D) or lung adenocarcinoma (image F). The immunostaining was performed on a Leica Biosystems BOND® RX instrument counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

  • Western blot - Anti-EGFRvIII antibody [EPR28380-83] (ab313646), expandable thumbnail

    Western blot - Anti-EGFRvIII antibody [EPR28380-83] (ab313646)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST
    This antibody does not recognize human wild-type EGFR. (A): anti-EGFRvIII antibody (ab313646) staining at 1/1000 dilution, (B): anti-EGFR antibody staining at 1/1000 dilution, (C): anti-GAPDH antibody [EPR16891] – Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

    All lanes: Western blot - Anti-EGFRvIII antibody [EPR28380-83] (ab313646) at 1/1000 dilution

    Lane 1: A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 3: EGFR knockout HeLa whole cell lysate at 20 µg

    Lane 4: Human placenta tissue lysate at 20 µg

    Secondary

    Lanes 1 - 3: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Lane 4: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Observed band size: 134 kDa

    Exposure time: 180s

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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