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Rabbit Monoclonal EGFR antibody. Carrier free. Suitable for IP, I-ELISA, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat, Recombinant full length protein - Human samples. Cited in 1 publication.

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Images

Immunoprecipitation - Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (AB174481), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (AB174481), expandable thumbnail
  • Western blot - Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (AB174481), expandable thumbnail
  • Western blot - Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (AB174481), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (AB174481), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

Constituents: PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPI-ELISAWBICC/IFFlow Cyt (Intra)IHC-P
Human
Tested
Expected
Tested
Tested
Tested
Tested
Mouse
Expected
Expected
Tested
Expected
Expected
Expected
Rat
Expected
Expected
Tested
Expected
Expected
Expected
Recombinant full length protein - Human
Not recommended
Tested
Not recommended
Not recommended
Not recommended
Not recommended

Tested
Tested

Species

Human

Dilution info

-

Notes

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

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Recombinant full length protein - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Recombinant full length protein - Human

Dilution info

1/2500

Notes

-

Expected
Expected

Species

Human, Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

-

Notes

Can be blocked with EGFR peptide (ab204282). This product yielded a strong signal in western blot using A431 (human squamous carcinoma) lysate which naturally overexpresses the EGFR protein. Western blot conditions may need to be optimised for cell lines and tissues that express lower levels of endogenous EGFR.

Species

Rat

Dilution info

-

Notes

Can be blocked with EGFR peptide (ab204282). This product yielded a strong signal in western blot using A431 (human squamous carcinoma) lysate which naturally overexpresses the EGFR protein. Western blot conditions may need to be optimised for cell lines and tissues that express lower levels of endogenous EGFR.

Species

Human

Dilution info

-

Notes

Can be blocked with EGFR peptide (ab204282). This product yielded a strong signal in western blot using A431 (human squamous carcinoma) lysate which naturally overexpresses the EGFR protein. Western blot conditions may need to be optimised for cell lines and tissues that express lower levels of endogenous EGFR.

Not recommended
Not recommended

Species

Recombinant full length protein - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Recombinant full length protein - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Recombinant full length protein - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

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

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Recombinant full length protein - Human

Dilution info

-

Notes

-

Associated Products

Select an associated product type

8 products for Alternative Version

7 products for Alternative Product

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:10805725, PubMed:27153536, PubMed:2790960, PubMed:35538033). Known ligands include EGF, TGFA/TGF-alpha, AREG, epigen/EPGN, BTC/betacellulin, epiregulin/EREG and HBEGF/heparin-binding EGF (PubMed:12297049, PubMed:15611079, PubMed:17909029, PubMed:20837704, PubMed:27153536, PubMed:2790960, PubMed:7679104, PubMed:8144591, PubMed:9419975). 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). Plays a role in mammalian pain signaling (long-lasting hypersensitivity) (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

Recommended products

Rabbit Monoclonal EGFR antibody. Carrier free. Suitable for IP, I-ELISA, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat, Recombinant full length protein - Human samples. Cited in 1 publication.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

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

Yes

Clone number

EP38Y

Purification technique

Affinity purification Protein A

Specificity

The immunogen for this product is a synthetic phospho-peptide corresponding to residues surrounding Tyr1068 of human EGFR. After screening, clone EP38Y was found to recognize total EGFR and is not specific to phosphorylated-Tyr1068 EGFR. This product yielded a strong signal in western blot using A431 (human squamous carcinoma) lysate which naturally overexpresses the EGFR protein. Western blot conditions may need to be optimised for cell lines and tissues that express lower levels of endogenous EGFR.
The mouse and rat recommendation is based on the WB results. This antibody may not be suitable for IHC with mouse or rat samples.

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

Notes

ab174481 is the carrier-free version of Anti-EGFR antibody [EP38Y] ab52894.

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

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Our Low endotoxin, azide-free formats have low endotoxin level (1 EU/mg, determined by the TAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

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:
Terms & Conditions.

8 product images

  • Immunoprecipitation - Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (ab174481), expandable thumbnail

    Immunoprecipitation - Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (ab174481)

    Anti-EGFR antibody [EP38Y] ab52894 (purified) at 1:20 dilution (0.5ug) immunoprecipitating EGFR in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate.

    Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug

    Lane 2 (+): Anti-EGFR antibody [EP38Y] ab52894 & HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate

    Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-EGFR antibody [EP38Y] ab52894 in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate

    For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used for detection at 1:1000 dilution.

    Blocking and diluting buffer: 5% NFDM/TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-EGFR antibody [EP38Y] ab52894).

    All lanes: Immunoprecipitation - Anti-EGFR antibody [EP38Y] (Anti-EGFR antibody [EP38Y] ab52894)

    Predicted band size: 134 kDa

    Observed band size: 175 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (ab174481), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (ab174481)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human cervical carcinoma tissue sections labeling EGFR with purified Anti-EGFR antibody [EP38Y] ab52894 at 1:100 dilution (0.95 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH9.0. Tissue was counterstained with Hematoxylin. Goat Anti-Rabbit IgG H&L (HRP) ab97051 Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1:500 dilution. PBS instead of the primary antibody was used as the negative control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-EGFR antibody [EP38Y] ab52894).

  • Western blot - Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (ab174481), expandable thumbnail

    Western blot - Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (ab174481)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-EGFR antibody [EP38Y] ab52894).

    False colour image of Western blot: Anti-EGFR antibody [EP38Y] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-EGFR antibody [EP38Y] ab52894 was shown to bind specifically to EGFR. A band was observed at 160 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in Egfr CRISPR-Cas9 edited cell line Human EGFR knockout HCT116 cell line ab281597 (CRISPR-Cas9 edited cell lysate ab282949). The band observed in the CRISPR-Cas9 edited lysate lane below 160 kDa is likely to represent a truncated form of EGFR. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and Egfr CRISPR-Cas9 edited HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.

    Lane 1: Wild-type HCT 116 cell lysate at 20 µg

    Lane 2: EGFR CRISPR-Cas9 edited HCT 116 cell lysate at 20 µg

    Lane 3: Caco-2 cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 134 kDa

    Observed band size: 160 kDa

  • Western blot - Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (ab174481), expandable thumbnail

    Western blot - Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (ab174481)

    This data was developed using the same antibody clone in a different buffer formulation (Anti-EGFR antibody [EP38Y] ab52894).

    Lanes 1 - 4: Merged signal (red and green). Green - Anti-EGFR antibody [EP38Y] ab52894 observed at 175 kDa. Red - loading control, Anti-Vinculin antibody [VIN-54] ab130007 observed at 125 kDa.

    Anti-EGFR antibody [EP38Y] ab52894 was shown to react with EGFR in wild-type HeLa. Loss of signal was observed when knockout cell line Human EGFR knockout HeLa cell line ab255385 (knockout cell lysate Human EGFR knockout HeLa cell lysate ab263845) was used. Wild-type and EGFR knockout samples were subjected to SDS-PAGE. Anti-EGFR antibody [EP38Y] ab52894 and Anti-Vinculin antibody [VIN-54] (Anti-Vinculin antibody [VIN-54] ab130007) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-EGFR antibody [EP38Y] (Anti-EGFR antibody [EP38Y] ab52894) at 1/1000 dilution

    Lane 1: A431 cell lysate at 20 µg

    Lane 2: MDA-MB-468 cell lysate at 20 µg

    Lane 3: Wild-type HeLa cell lysate at 20 µg

    Lane 4: EGFR knockout HeLa cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution

    Performed under reducing conditions.

    Predicted band size: 134 kDa

    Observed band size: 124 kDa, 134 kDa

    Lanes 1 - 4: Merged signal (red and green). Green - Anti-EGFR antibody [EP38Y] ab52894 observed at 175 kDa. Red - loading control, Anti-Vinculin antibody [VIN-54] ab130007 observed at 125 kDa.

    Anti-EGFR antibody [EP38Y] ab52894 was shown to react with EGFR in wild-type HeLa. Loss of signal was observed when knockout cell line Human EGFR knockout HeLa cell line ab255385 (knockout cell lysate Human EGFR knockout HeLa cell lysate ab263845) was used. Wild-type and EGFR knockout samples were subjected to SDS-PAGE. Anti-EGFR antibody [EP38Y] ab52894 and Anti-Vinculin antibody [VIN-54] (Anti-Vinculin antibody [VIN-54] ab130007) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Flow Cytometry (Intracellular) - Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (ab174481), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (ab174481)

    Intracellular Flow Cytometry analysis of A431 (Human epidermoid carcinoma epithelial cell) cells labelling EGFR with purified Anti-EGFR antibody [EP38Y] ab52894. Cells were fixed with 4% Paraformaldehyde (10min) and permeabilised with 90% methanol for 30min. Then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by Anti-EGFR antibody [EP38Y] ab52894 at 1/20 dilution (red) for 30 min. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-EGFR antibody [EP38Y] ab52894).

  • Immunocytochemistry/ Immunofluorescence - Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (ab174481), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (ab174481)

    Immunocytochemistry/ Immunofluorescence analysis of A431 (Human epidermoid carcinoma epithelial cell) cells labeling EGFR with Purified Anti-EGFR antibody [EP38Y] ab52894 at 1:250 dilution (0.4μg/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) 1:200 (2.5 μg/ml). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-EGFR antibody [EP38Y] ab52894).

  • Indirect ELISA - Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (ab174481), expandable thumbnail

    Indirect ELISA - Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (ab174481)

    ELISA analysis of Human EGFR recombinant protein at 1000 ng/ml with Anti-EGFR antibody [EP38Y] ab52894. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.

  • OI-RD Scanning - Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (ab174481), expandable thumbnail

    OI-RD Scanning - Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (ab174481)

    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.

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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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