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Phospho EGFR antibody pY1068 [EP774Y] ab40815 is a rabbit monoclonal antibody that is used in EGFR western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human and mouse samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone EP774Y has been tried and trusted by researchers since 2006 and is cited in >190 publications
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- One antibody for all your EGFR staining, use in EGFR western blotting, IHC, immunofluorescence and flow cytometry

New 20 ul size available


Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR (phospho Y1068) antibody [EP774Y] (AB40815), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR (phospho Y1068) antibody [EP774Y] (AB40815), expandable thumbnail
  • Western blot - Anti-EGFR (phospho Y1068) antibody [EP774Y] (AB40815), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-EGFR (phospho Y1068) antibody [EP774Y] (AB40815), expandable thumbnail
  • Western blot - Anti-EGFR (phospho Y1068) antibody [EP774Y] (AB40815), expandable thumbnail

Publications

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
DotWBICC/IFFlow Cyt (Intra)IHC-P
Human
Tested
Tested
Tested
Tested
Tested
Mouse
Expected
Tested
Expected
Expected
Expected

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

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

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/500 - 1/5000

Notes

-

Species

Human

Dilution info

1/500 - 1/5000

Notes

-

Tested
Tested

Species

Human

Dilution info

1/250 - 1/500

Notes

-

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/800

Notes

-

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/250 - 1/500

Notes

The mouse recommendation is based on the WB results. We do not guarantee IHC-P for mouse.

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

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

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Target data

Function

Receptor tyrosine kinase binding ligands of the EGF family and activating several signaling cascades to convert extracellular cues into appropriate cellular responses (PubMed:2790960, PubMed:10805725, PubMed:27153536). Known ligands include EGF, TGFA/TGF-alpha, AREG, epigen/EPGN, BTC/betacellulin, epiregulin/EREG and HBEGF/heparin-binding EGF (PubMed:2790960, PubMed:7679104, PubMed:8144591, PubMed:9419975, PubMed:15611079, PubMed:12297049, PubMed:27153536, PubMed:20837704, PubMed:17909029). Ligand binding triggers receptor homo- and/or heterodimerization and autophosphorylation on key cytoplasmic residues. The phosphorylated receptor recruits adapter proteins like GRB2 which in turn activates complex downstream signaling cascades. Activates at least 4 major downstream signaling cascades including the RAS-RAF-MEK-ERK, PI3 kinase-AKT, PLCgamma-PKC and STATs modules (PubMed:27153536). May also activate the NF-kappa-B signaling cascade (PubMed:11116146). Also directly phosphorylates other proteins like RGS16, activating its GTPase activity and probably coupling the EGF receptor signaling to the G protein-coupled receptor signaling (PubMed:11602604). Also phosphorylates MUC1 and increases its interaction with SRC and CTNNB1/beta-catenin (PubMed:11483589). Positively regulates cell migration via interaction with CCDC88A/GIV which retains EGFR at the cell membrane following ligand stimulation, promoting EGFR signaling which triggers cell migration (PubMed:20462955). Plays a role in enhancing learning and memory performance (By similarity).Isoform 2 may act as an antagonist of EGF action.(Microbial infection) Acts as a receptor for hepatitis C virus (HCV) in hepatocytes and facilitates its cell entry. Mediates HCV entry by promoting the formation of the CD81-CLDN1 receptor complexes that are essential for HCV entry and by enhancing membrane fusion of cells expressing HCV envelope glycoproteins.

Alternative names

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

Recommended products

Phospho EGFR antibody pY1068 [EP774Y] ab40815 is a rabbit monoclonal antibody that is used in EGFR western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human and mouse samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone EP774Y has been tried and trusted by researchers since 2006 and is cited in >190 publications
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- One antibody for all your EGFR staining, use in EGFR western blotting, IHC, immunofluorescence and flow cytometry

New 20 ul size available

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

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

EP774Y

Purification technique

Affinity purification Protein A

Specificity

Recognises EGFR phosphorylated on Tyrosine 1068 of the mature human isoform 1 (corresponding to Y1092 from the precursor form P00533-1/p170)

The mouse recommendation is based on the WB results. We do not guarantee IHC-P for mouse.

Dissociation constant

3.6 x 10-11 M

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 or Epidermal Growth Factor Receptor is a transmembrane glycoprotein that acts as a receptor for members of the epidermal growth factor family. Known alternatively as ErbB1 or HER1 this receptor has an approximate molecular weight of 170 kDa. EGFR is expressed in various cell types notably on epithelial cells and can influence multiple cellular processes through its kinase activity. It participates in the regulation of cell growth multiplication and survival by activating its kinase domain upon ligand binding.

Biological function summary

The EGFR protein plays an important role in cellular communication and signaling processes. EGFR pairs with other receptor family members to form active dimers or even higher-order complexes which in turn initiate intracellular signaling cascades. Through these complexes EGFR influences many processes including cell differentiation and repair. This function of EGFR makes it an integral part of mammalian biology affecting how cells respond to their environment by mediating changes in gene expression.

Pathways

EGFR is a central player in the MAPK and PI3K/Akt signaling pathways. Alongside other protein partners like KRAS and PI3 kinase it contributes to transmitting signals from the cell surface to the nucleus affecting gene transcription and cell behavior. These pathways are important for normal cell growth and division and aberrations in these pathways can lead to excessive or insufficient cell proliferation.

Associated diseases and disorders

EGFR is pertinent to cancer biology including non-small cell lung cancer and glioblastoma where mutations or overexpression of the receptor frequently occur. It connects to proteins such as PTEN and BRAF which influence tumor progression and response to targeted therapies. EGFR's involvement in these disorders highlights its significance as a therapeutic target since it can be manipulated to alter disease progression.

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:
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14 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815), expandable thumbnail
    This image is courtesy of an anonymous customer review.

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815)

    Formaldehyde-fixed, paraffin-embedded human prostate cancer tissue stained for EGFR (phospho Y1068) using ab40815 (unpurified) at 1/200 dilution in immunohistochemical analysis, followed by Goat anti Rabbit IgG (Biotin).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast cancer tissue sections labeling EGFR with purified ab40815 at 1:500 dilution (1.75 µg/ml). Heat mediated antigen retrieval was performed using heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Western blot - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815), expandable thumbnail

    Western blot - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815)

    This antibody detects high background.

    All lanes: Western blot - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815) at 1/1000 dilution

    Lane 1: A431 (Human epidermoid carcinoma epithelial cell) whole cell lysates at 15 µg

    Lane 2: A431 (Human epidermoid carcinoma epithelial cell) treated with 10 ng/ml EGF for 30 minutes whole cell lysates at 15 µg

    Lane 3: A431 (Human epidermoid carcinoma epithelial cell) treated with 10 ng/ml EGF for 30 minutes. Then the membrane was incubated with phosphatase at 15 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 134 kDa

    Observed band size: 175 kDa

  • Flow Cytometry (Intracellular) - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815)

    Intracellular Flow Cytometry analysis of A431 (Human epidermoid carcinoma epithelial cell) treated with 200 ng/ml EGF for 15 minutes cells labeling EGFR with purified ab40815 at 1/800 dilution (1 μg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). Untreated A431 cells (Green).

  • Western blot - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815), expandable thumbnail

    Western blot - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815)

    This antibody detects high background.

    All lanes: Western blot - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815) at 1/1000 dilution

    Lane 1: A431 (Human epidermoid carcinoma epithelial cell) whole cell lysates at 15 µg

    Lane 2: A431 (Human epidermoid carcinoma epithelial cell) treated with 10 ng/ml EGF for 30 minutes whole cell lysates at 15 µg

    Lane 3: A431 (Human epidermoid carcinoma epithelial cell) treated with 10 ng/ml EGF for 30 minutes. Then the membrane was incubated with phosphatase at 15 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 134 kDa

    Observed band size: 175 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815)

    Immunocytochemistry/ Immunofluorescence analysis of A431 (Human epidermoid carcinoma epithelial cell) treated with 100 ng/ml EGF for 10 minutes cells labeling EGFR with purified ab40815 at 1:500 dilution (1.8 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815)

    WD-PBEC cultures were infected with RSV clinical isolate BT2a and stained for EGFR (red) and RSV F (RSV F, green) expression.

    For WD-PBECs, pediatric bronchial epithelial cells (PBEC) were obtained, via written parental consent, from bronchial brushings of children undergoing elective surgery at the Royal Belfast Hospital for Sick Children, and the procedures were approved by the Office of Research Ethics Committees Northern Ireland. PBEC were expanded in collagen-coated flasks using airway epithelial cell media and supplements (Lonza), then seeded onto transwell inserts (Corning), and then air-liquid interface (ALI) cultures were initiated and maintained 21 days in order to establish well-differentiated (WD)-PBECs. Paraformaldehyde-fixed and permeabilized WD-PBEC were stained for RSV F protein expression and were stained with anti-phospho-(p)-EGFR (Abcam, ab40815, unpurified). WD-PBEC cultures were infected with RSV subgroup A clinical isolate BT2a. Fluorescent images were obtained with a SP5 confocal DMI 6000 inverted microscope (Leica).

  • Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815)

    Immunofluorescent staining of untreated (C) and Phosphatase- treated (D) A431 cells using ab40815 (unpurified).

  • Dot Blot - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815), expandable thumbnail

    Dot Blot - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815)

    Dot blot analysis of EGFR (pY1068) peptide (Lane 1), SMAD5 (unmodified) peptide labelling EGFR (pY1068) with ab40815 (unpurified) at a dilution of 1/1000. Peroxidase conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody at a dilution of 1/2500.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815)

    ab40815 (unpurified) showing positive staining in Papillary carcinoma of thyroid gland tissue.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815)

    ab40815 (unpurified) showing positive staining in Glioma tissue.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815)

    ab40815 (unpurified) showing positive staining in Cervical carcinoma tissue.

  • OI-RD Scanning - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815), expandable thumbnail

    OI-RD Scanning - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815)

    We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
    Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

  • Western blot - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815), expandable thumbnail

    Western blot - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815)

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

    Lanes 1 - 3: Western blot - Anti-EGFR (phospho Y1068) antibody [EP774Y] (ab40815) at 1/1000 dilution

    Lanes 1 - 3: Western blot - Anti-EGFR (phospho Y1068) antibody [EP774Y] - BSA and Azide free (Anti-EGFR (phospho Y1068) antibody [EP774Y] - BSA and Azide free ab182618) at 1/1000 dilution

    Lane 1: C2C12 (Mouse myoblasts myoblast) whole cell lysate at 20 µg

    Lane 2: C2C12 (Mouse myoblasts myoblast) treated with 100 ng/ml EGF for 24 hours whole cell lysate at 20 µg

    Lane 3: C2C12 (Mouse myoblasts myoblast) treated with 100 ng/ml EGF for 24hours whole cell lysate. Then the membrane was incubated with phosphatase. at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Exposure time: 30s

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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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