For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome

Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
United States
Your country/region is currently set to:

If incorrect, please enter your country/region into the box below, to view site information related to your country/region.

Call (888) 77-ABCAM (22226) or contact us
Need help? Contact us

  • My account
  • Sign out
Sign in or Register with us

Welcome

Sign in or

Don't have an account?

Register with us
My basket
Quick order
Abcam homepage

  • Research Products
    By product type
    Primary antibodies
    Secondary antibodies
    ELISA and Matched Antibody Pair Kits
    Cell and tissue imaging tools
    Cellular and biochemical assays
    Proteins and Peptides
    By product type
    Proteomics tools
    Agonists, activators, antagonists and inhibitors
    Cell lines and Lysates
    Multiplex miRNA assays
    Multiplex Assays
    By research area
    Cancer
    Cardiovascular
    Cell Biology
    Epigenetics
    Metabolism
    Developmental Biology
    By research area
    Immunology
    Microbiology
    Neuroscience
    Signal Transduction
    Stem Cells
  • Customized Products & Partnerships
    Customized Products & Partnerships

    Customized products and commercial partnerships to accelerate your diagnostic and therapeutic programs.

    Customized products

    Partner with us

  • Support
    Support hub

    Access advice and support for any research roadblock

    View support hub

    Protocols

    Your experiments laid out step by step

    View protocols

  • Events
    • Conference calendar
    • Cancer
    • Cardiovascular
    • Epigenetics & Nuclear signaling
    • Immunology
    • Neuroscience
    • Stem cells
    • Tradeshows
    • Scientific webinars
    Keep up to date with the latest events

    Full event breakdown with abstracts, speakers, registration and more

    View global event calendar

  • Pathways
    Cell signalling pathways

    View all pathways

    View all interactive pathways

  1. Link

    products/primary-antibodies/egfr-antibody-ep38y-ab52894.pdf

  1. Send me a copy of this email
    I agree to the terms and conditions.
Signal Transduction Protein Phosphorylation Tyrosine Kinases Receptor Tyrosine Kinases
Share by email
Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-EGFR antibody [EP38Y] (ab52894)

  • Datasheet
  • SDS
Reviews (22)Q&A (1)References (244)

Product price, shipping and contact information

Currently unavailable

Sorry, we can't display this right now.

Please contact us to place your order, or try again later.

 

Loading size & price…

 

Shipping and order information

Shipping info

Promotion Information

Abpromise

Guaranteed product quality, expert customer support.

Find out more.

Western blot - Anti-EGFR antibody [EP38Y] (ab52894)
  • Western blot - Anti-EGFR antibody [EP38Y] (ab52894)
  • Western blot - Anti-EGFR antibody [EP38Y] (ab52894)
  • Immunocytochemistry/ Immunofluorescence - Anti-EGFR antibody [EP38Y] (ab52894)
  • Indirect ELISA - Anti-EGFR antibody [EP38Y] (ab52894)
  • Western blot - Anti-EGFR antibody [EP38Y] (ab52894)
  • Western blot - Anti-EGFR antibody [EP38Y] (ab52894)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR antibody [EP38Y] (ab52894)
  • Immunocytochemistry/ Immunofluorescence - Anti-EGFR antibody [EP38Y] (ab52894)
  • Flow Cytometry (Intracellular) - Anti-EGFR antibody [EP38Y] (ab52894)
  • Immunoprecipitation - Anti-EGFR antibody [EP38Y] (ab52894)
  • OI-RD Scanning - Anti-EGFR antibody [EP38Y] (ab52894)
  • Anti-EGFR antibody [EP38Y] (ab52894)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP38Y] to EGFR
  • Suitable for: WB, IP, IHC-P, ICC/IF, Indirect ELISA, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free HRP PE

You may also be interested in

Primary
Product image
Anti-ErbB2 / HER2 (phospho Y877) antibody [EP2324Y] (ab108371)
Primary
Product image
Alexa Fluor® 488 Anti-EGFR antibody [EP38Y] (ab193244)
Knockout
Product image
Human EGFR knockout HeLa cell line (ab255385)

View more associated products

Overview

  • Product name

    Anti-EGFR antibody [EP38Y]
    See all EGFR primary antibodies
  • Description

    Rabbit monoclonal [EP38Y] to EGFR
  • Host species

    Rabbit
  • 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.

  • Tested applications

    Suitable for: WB, IP, IHC-P, ICC/IF, Indirect ELISA, Flow Cyt (Intra)more details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
    (Peptide available as ab204282)

  • Positive control

    • ICC/IF: A431 cells. WB: HeLa, Caco-2 and A431 cell lysate; Rat liver and skin lysate; Mouse lung and skin lysate IP: HeLa whole cell lysate (ab150035). Flow Cyt (intra): A431 cells. IHC-P: Human cervical carcinoma
  • General notes

    Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).

    See other anti-rabbit secondary antibodies that can be used with this antibody.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Dissociation constant (KD)

    KD = 1.90 x 10 -11 M
    Learn more about KD
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP38Y
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Protein Phosphorylation
    • Tyrosine Kinases
    • Receptor Tyrosine Kinases
    • Signal Transduction
    • Growth Factors/Hormones
    • EGF
    • Cell Biology
    • Cell Cycle
    • Cell differentiation
    • Microbiology
    • Organism
    • Virus
    • RNA Virus
    • ssRNA positive strand virus
    • SARS Coronavirus
    • Cancer
    • Growth factors
    • EGF
    • Cancer
    • Signal transduction
    • Protein phosphorylation
    • Tyrosine kinases
    • Receptor tyrosine kinases
    • Cancer
    • Oncoproteins/suppressors
    • Oncoproteins
    • Growth factor receptors
    • Cancer
    • Tumor biomarkers
    • Receptors
    • Signal Transduction
    • Antibodies
    • egfr
    • Neuroscience
    • Development
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (ab174481)
    • Alexa Fluor® 647 Anti-EGFR antibody [EP38Y] (ab192982)
    • Alexa Fluor® 488 Anti-EGFR antibody [EP38Y] (ab193244)
    • HRP Anti-EGFR antibody [EP38Y] (ab193602)
    • Alexa Fluor® 594 Anti-EGFR antibody [EP38Y] (ab207870)
    • PE Anti-EGFR antibody [EP38Y] (ab208753)
    • Anti-EGFR antibody [EP38Y] - BSA and Azide free (ab272293)
    • Alexa Fluor® 555 Anti-EGFR antibody [EP38Y] (ab274892)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Immunizing Peptide (Blocking)

    • EGFR peptide (ab204282)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human EGFR knockout HeLa cell line (ab255385)
  • KO cell lysates

    • Human EGFR knockout HeLa cell lysate (ab263845)
  • Positive Controls

    • HeLa whole cell lysate (ab29545)
    • HeLa membrane extract lysate (ab29547)
  • Recombinant Protein

    • Recombinant human EGFR protein (Active) (ab155639)
    • Recombinant human EGFR protein (Fc Chimera) (ab84002)
  • Related Products

    • Human EGFR ELISA Kit (ERBB) (ab100505)
    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)
    • Anti-EGFR (phospho Y1086) antibody [Y39] (ab32086)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab52894 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB (12)
1/1000 - 1/10000. Detects a band of approximately 175 kDa (predicted molecular weight: 134 kDa).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

IP
1/20.
IHC-P (5)
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

The mouse and rat recommendation is based on the WB results. This antibody may not be suitable for IHC with mouse or rat samples.

ICC/IF (3)
1/250 - 1/500.
Indirect ELISA
1/2500.
Flow Cyt (Intra)
1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Notes
WB
1/1000 - 1/10000. Detects a band of approximately 175 kDa (predicted molecular weight: 134 kDa).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

IP
1/20.
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

The mouse and rat recommendation is based on the WB results. This antibody may not be suitable for IHC with mouse or rat samples.

ICC/IF
1/250 - 1/500.
Indirect ELISA
1/2500.
Flow Cyt (Intra)
1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Function

    Receptor tyrosine kinase binding ligands of the EGF family and activating several signaling cascades to convert extracellular cues into appropriate cellular responses. Known ligands include EGF, TGFA/TGF-alpha, amphiregulin, epigen/EPGN, BTC/betacellulin, epiregulin/EREG and HBEGF/heparin-binding EGF. 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. May also activate the NF-kappa-B signaling cascade. 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. Also phosphorylates MUC1 and increases its interaction with SRC and CTNNB1/beta-catenin.
    Isoform 2 may act as an antagonist of EGF action.
  • Tissue specificity

    Ubiquitously expressed. Isoform 2 is also expressed in ovarian cancers.
  • Involvement in disease

    Lung cancer
    Inflammatory skin and bowel disease, neonatal, 2
  • Sequence similarities

    Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications

    Phosphorylation at Ser-695 is partial and occurs only if Thr-693 is phosphorylated. Phosphorylation at Thr-678 and Thr-693 by PRKD1 inhibits EGF-induced MAPK8/JNK1 activation. Dephosphorylation by PTPRJ prevents endocytosis and stabilizes the receptor at the plasma membrane. Autophosphorylation at Tyr-1197 is stimulated by methylation at Arg-1199 and enhances interaction with PTPN6. Autophosphorylation at Tyr-1092 and/or Tyr-1110 recruits STAT3. Dephosphorylated by PTPN1 and PTPN2.
    Monoubiquitinated and polyubiquitinated upon EGF stimulation; which does not affect tyrosine kinase activity or signaling capacity but may play a role in lysosomal targeting. Polyubiquitin linkage is mainly through 'Lys-63', but linkage through 'Lys-48', 'Lys-11' and 'Lys-29' also occurs. Deubiquitination by OTUD7B prevents degradation. Ubiquitinated by RNF115 and RNF126.
    Methylated. Methylation at Arg-1199 by PRMT5 stimulates phosphorylation at Tyr-1197.
  • Cellular localization

    Secreted and Cell membrane. Endoplasmic reticulum membrane. Golgi apparatus membrane. Nucleus membrane. Endosome. Endosome membrane. Nucleus. In response to EGF, translocated from the cell membrane to the nucleus via Golgi and ER. Endocytosed upon activation by ligand. Colocalized with GPER1 in the nucleus of estrogen agonist-induced cancer-associated fibroblasts (CAF).
  • Target information above from: UniProt accession P00533 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 1956 Human
    • Entrez Gene: 13649 Mouse
    • Entrez Gene: 24329 Rat
    • Omim: 131550 Human
    • SwissProt: P00533 Human
    • SwissProt: Q01279 Mouse
    • Unigene: 488293 Human
    • Unigene: 605083 Human
    • Unigene: 420648 Mouse
    • Unigene: 439882 Mouse
    • Unigene: 8534 Mouse
    see all
  • Alternative names

    • Avian erythroblastic leukemia viral (v erb b) oncogene homolog antibody
    • Cell growth inhibiting protein 40 antibody
    • Cell proliferation inducing protein 61 antibody
    • EGF R antibody
    • EGFR antibody
    • EGFR_HUMAN antibody
    • Epidermal growth factor receptor (avian erythroblastic leukemia viral (v erb b) oncogene homolog) antibody
    • Epidermal growth factor receptor (erythroblastic leukemia viral (v erb b) oncogene homolog avian) antibody
    • Epidermal growth factor receptor antibody
    • erb-b2 receptor tyrosine kinase 1 antibody
    • ERBB antibody
    • ERBB1 antibody
    • Errp antibody
    • HER1 antibody
    • mENA antibody
    • NISBD2 antibody
    • Oncogen ERBB antibody
    • PIG61 antibody
    • Proto-oncogene c-ErbB-1 antibody
    • Receptor tyrosine protein kinase ErbB 1 antibody
    • Receptor tyrosine-protein kinase ErbB-1 antibody
    • SA7 antibody
    • Species antigen 7 antibody
    • Urogastrone antibody
    • v-erb-b Avian erythroblastic leukemia viral oncogen homolog antibody
    • wa2 antibody
    • Wa5 antibody
    see all

Images

  • Western blot - Anti-EGFR antibody [EP38Y] (ab52894)
    Western blot - Anti-EGFR antibody [EP38Y] (ab52894)
    All lanes : Anti-EGFR antibody [EP38Y] (ab52894) at 1/10000 dilution (purified)

    Lane 1 : Rat liver lysates
    Lane 2 : Mouse lung lysates

    Lysates/proteins at 15 µg per lane.

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

    Predicted band size: 134 kDa
    Observed band size: 175 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer: 5% NFDM/TBST

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

    Lane 1 : A431 cell lysate
    Lane 2 : MDA-MB-468 cell lysate
    Lane 3 : Wild-type HeLa cell lysate
    Lane 4 : EGFR knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 134 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab52894 observed at 175 kDa. Red - loading control, ab130007 observed at 125 kDa.  

     ab52894 was shown to react with EGFR in wild-type HeLa. Loss of signal was observed when knockout cell line ab255385 (knockout cell lysate ab263845) was used. Wild-type and EGFR knockout samples were subjected to SDS-PAGE. ab52894 and 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 (ab216773) and 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.

     

  • Western blot - Anti-EGFR antibody [EP38Y] (ab52894)
    Western blot - Anti-EGFR antibody [EP38Y] (ab52894)
    Anti-EGFR antibody [EP38Y] (ab52894) at 1/2000 dilution (purified) + HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 134 kDa
    Observed band size: 175 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer: 5% NFDM/TBST

  • Immunocytochemistry/ Immunofluorescence - Anti-EGFR antibody [EP38Y] (ab52894)
    Immunocytochemistry/ Immunofluorescence - Anti-EGFR antibody [EP38Y] (ab52894)

    Unpurified ab52894 stained A431 (Human epidermoid carcinoma cell line) cells.

    The cells were fixed in 100% methanol for 5 mins at -20°C and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour at room temperature to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab52894 at 1 in 500) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1 hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43 µM for 1 hour at room temperature.

  • Indirect ELISA - Anti-EGFR antibody [EP38Y] (ab52894)
    Indirect ELISA - Anti-EGFR antibody [EP38Y] (ab52894)

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

  • Western blot - Anti-EGFR antibody [EP38Y] (ab52894)
    Western blot - Anti-EGFR antibody [EP38Y] (ab52894)
    Different batches of ab52894 were tested on HeLa (Human cervix adenocarcinoma epithelial cell) lysate at 1.0 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 175 kDa.
  • Western blot - Anti-EGFR antibody [EP38Y] (ab52894)
    Western blot - Anti-EGFR antibody [EP38Y] (ab52894)
    All lanes : Anti-EGFR antibody [EP38Y] (ab52894) at 1/1000 dilution (unpurified)

    Lane 1 : Caco-2 (Human colorectal adenocarcinoma cell line) cell lysate
    Lane 2 : A431 (Human epidermoid carcinoma cell line) cell lysate
    Lane 3 : Mouse skin cell lysate
    Lane 4 : Rat skin cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 134 kDa



    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 mins before being transferred onto a Nitrocellulose membrane at 30V for 70 mins. The membrane was then blocked for an hour before being incubated with unpurified ab52894 overnight at 4°C in the presence of loading control ab18058 (Mouse monoclonal [SPM227] to Vinculin diluted 1:10000). Antibody binding was detected using IR-labeled goat anti-Rabbit Ab at a 1:10,000 dilution for one hour at room temperature before imaging.

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR antibody [EP38Y] (ab52894)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR antibody [EP38Y] (ab52894)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue sections labeling EGFR with purified ab52894 at 1:100 dilution (0.95 μg/ml).

    Heat mediated antigen retrieval was performed using EDTA buffer, pH 9.0. Tissue was counterstained with hematoxylin. 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-EGFR antibody [EP38Y] (ab52894)
    Immunocytochemistry/ Immunofluorescence - Anti-EGFR antibody [EP38Y] (ab52894)

    Immunocytochemistry/ Immunofluorescence analysis of A431 (Human epidermoid carcinoma epithelial cell) cells labeling EGFR with purified ab52894 at 1:250 dilution (0.4 μg/ml).

    Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). 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.

  • Flow Cytometry (Intracellular) - Anti-EGFR antibody [EP38Y] (ab52894)
    Flow Cytometry (Intracellular) - Anti-EGFR antibody [EP38Y] (ab52894)

    Intracellular Flow Cytometry analysis of A431 (Human epidermoid carcinoma epithelial cell) cells labeling EGFR with purified ab52894. 

    Cells were fixed with 4% paraformaldehyde (10 mins) and permeabilized with 90% methanol for 30 mins. Then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by ab52894 at 1/20 dilution (red) for 30 mins. A Goat anti rabbit IgG (Alexa Fluorr® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). 

     

     

  • Immunoprecipitation - Anti-EGFR antibody [EP38Y] (ab52894)
    Immunoprecipitation - Anti-EGFR antibody [EP38Y] (ab52894)

    ab52894 (purified) at 1:20 dilution (0.5 µg) immunoprecipitating EGFR in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate.

    Lane 1 (input): HeLa whole cell lysate 10 µg

    Lane 2 (+): ab52894 in HeLa whole cell lysate

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52894 in HeLa whole cell lysate

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

    Blocking and diluting buffer: 5% NFDM/TBST.

  • OI-RD Scanning - Anti-EGFR antibody [EP38Y] (ab52894)
    OI-RD Scanning - Anti-EGFR antibody [EP38Y] (ab52894)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • Anti-EGFR antibody [EP38Y] (ab52894)
    Anti-EGFR antibody [EP38Y] (ab52894)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (244)

Publishing research using ab52894? Please let us know so that we can cite the reference in this datasheet.

ab52894 has been referenced in 244 publications.

  • Pereira PMR  et al. Immuno-PET Detects Changes in Multi-RTK Tumor Cell Expression Levels in Response to Targeted Kinase Inhibition. J Nucl Med 62:366-371 (2021). PubMed: 32646879
  • Chen Y  et al. CDKN2A (p16INK4A) affects the anti-tumor effect of CDK inhibitor in somatotroph adenomas. Int J Mol Med 47:500-510 (2021). PubMed: 33416096
  • Wang S  et al. AXL is a candidate receptor for SARS-CoV-2 that promotes infection of pulmonary and bronchial epithelial cells. Cell Res 31:126-140 (2021). PubMed: 33420426
  • Cao X  et al. Alt-RPL36 downregulates the PI3K-AKT-mTOR signaling pathway by interacting with TMEM24. Nat Commun 12:508 (2021). PubMed: 33479206
  • Zhang A  et al. TNK2 promoted esophageal cancer progression via activating egfr-akt signaling. J Clin Lab Anal 35:e23700 (2021). PubMed: 33484472
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-10 of 23 Abreviews or Q&A

Western blot abreview for Anti-EGFR antibody [EP38Y]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (non small cell lung cancer)
Gel Running Conditions
Reduced Denaturing (4-15%)
Loading amount
20 µg
Treatment
siRNA treated
Specification
non small cell lung cancer
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 07 2014

Western blot abreview for Anti-EGFR antibody [EP38Y]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Rat Tissue lysate - whole (Liver)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (10)
Loading amount
20 µg
Specification
Liver
Blocking step
BSA as blocking agent for 30 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 30 2022

Western blot abreview for Anti-EGFR antibody [EP38Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa cells)
Gel Running Conditions
Reduced Denaturing (Bolt 4-12% Bis Tris Plus)
Loading amount
10 µg
Specification
HeLa cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 15 2022

Immunocytochemistry/ Immunofluorescence abreview for Anti-EGFR antibody [EP38Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa cells)
Permeabilization
Yes - 0.5% Triton X-100 in PBS, 5min, RT
Specification
HeLa cells
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Mr. Carlos Pardo-Pastor

Verified customer

Submitted Mar 14 2022

Western blot abreview for Anti-EGFR antibody [EP38Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (Liver)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (10)
Loading amount
30 µg
Treatment
200µM Palmitate
Specification
Liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 05 2021

Western blot abreview for Anti-EGFR antibody [EP38Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Purified protein (Melanoma Cells)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
30 µg
Specification
Melanoma Cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Aug 12 2020

Western blot abreview for Anti-EGFR antibody [EP38Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Liver)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (10)
Loading amount
20 µg
Treatment
200µM Palmitate
Specification
Liver
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jul 06 2020

Western blot abreview for Anti-EGFR antibody [EP38Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (U87 cells)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (4-20% gel)
Loading amount
15 µg
Treatment
bevacizumab 2mg/ml for 24hrs, 48hrs
Specification
U87 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 11 2020

Western blot abreview for Anti-EGFR antibody [EP38Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Rat Cell lysate - whole cell (Hippocampal culture)
Gel Running Conditions
Reduced Denaturing (4-15% gradient)
Loading amount
15 µg
Specification
Hippocampal culture
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 27 2020

Western blot abreview for Anti-EGFR antibody [EP38Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (Liver)
Gel Running Conditions
Reduced Denaturing (4-15% gradient)
Loading amount
40 µg
Specification
Liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 23 2020

1-10 of 23 Abreviews or Q&A

  •  Previous
  • 1
  • 2
  • 3
  • Next 

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Get resources and offers direct to your inbox Sign up
A-Z by research area
  • Cancer
  • Cardiovascular
  • Cell biology
  • Developmental biology
  • Epigenetics & Nuclear signaling
  • Immunology
  • Metabolism
  • Microbiology
  • Neuroscience
  • Signal transduction
  • Stem cells
A-Z by product type
  • Primary antibodies
  • Secondary antibodies
  • Biochemicals
  • Isotype controls
  • Flow cytometry multi-color selector
  • Kits
  • Loading controls
  • Lysates
  • Peptides
  • Proteins
  • Slides
  • Tags and cell markers
  • Tools & Reagents
Help & support
  • Support
  • Make an Inquiry
  • Protocols & troubleshooting
  • Placing an order
  • RabMAb products
  • Biochemical product FAQs
  • Training
  • Browse by Target
Company
  • Corporate site
  • Investor relations
  • Company news
  • Careers
  • About us
  • Blog
Events
  • Tradeshows
  • Conferences
International websites
  • abcam.cn
  • abcam.co.jp

Join with us

  • LinkedIn
  • facebook
  • Twitter
  • YouTube
  • Terms of sale
  • Website terms of use
  • Cookie policy
  • Privacy policy
  • Legal
  • Modern slavery statement
© 1998-2023 Abcam plc. All rights reserved.