概述

  • 产品名称Anti-EGFR抗体[EP38Y]
    参阅全部 EGFR 一抗
  • 描述
    兔单克隆抗体[EP38Y] to EGFR
  • 特异性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
  • 经测试应用适用于: WB, IP, Flow Cyt, IHC-P, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human EGFR. Synthetic phospho-peptide corresponding to residues surrounding Tyr1068 of mature human EGFR.
    Database link: P00533

  • 阳性对照
    • A431, HeLa cell lysate or human lymph node metastasis tissue.
  • 常规说明

    A trial size is available to purchase for this antibody.

    Alternative versions available:
    Anti-EGFR antibody (Alexa Fluor® 488) [EP38Y] (ab193244)
    Anti-EGFR antibody (Alexa Fluor® 647) [EP38Y] (ab192982)
    Anti-EGFR antibody (HRP) [EP38Y] (ab193602)
    Anti-EGFR antibody (Alexa Fluor® 594) [EP38Y] (ab207870)
    Anti-EGFR antibody (Phycoerythrin) [EP38Y] (ab208753)

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

性能

应用

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.

应用 Ab评论 说明
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 1/50.
Flow Cyt 1/100.

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

IHC-P Use at an assay dependent concentration.
ICC/IF 1/250 - 1/500.

靶标

  • 功能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.
  • 组织特异性Ubiquitously expressed. Isoform 2 is also expressed in ovarian cancers.
  • 疾病相关Lung cancer
    Inflammatory skin and bowel disease, neonatal, 2
  • 序列相似性Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.
    Contains 1 protein kinase domain.
  • 翻译后修饰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.
  • 细胞定位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).
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

Anti-EGFR antibody [EP38Y] 图像

  • ab52894 stained A431 cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab52894 at 1in500) 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 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

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

    Lane 1 : Caco-2 cell lysate
    Lane 2 : A431 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 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with 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-labelled 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.

  • ab52894 showing positive staining in Endometrial carcinoma tissue.

  • Overlay histogram showing A431 cells stained with ab52894 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52894, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti-rabbit DyLight® 488 (IgG H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

  • Anti-EGFR antibody [EP38Y] (ab52894) at 1/100000 dilution + HeLa cell lysate at 10 µg

    Secondary
    Goat anti-Rabbit HRP labeled. at 1/2000 dilution

    Predicted band size : 134 kDa
    Observed band size : 175 kDa (why is the actual band size different from the predicted?)
  • ab52894 showing positive staining in Cervical carcinoma tissue.

  • Anti-EGFR antibody [EP38Y] (ab52894) at 1/500 dilution + Murine macrophage lysates at 80 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 134 kDa
    Additional bands at : 170 kDa (possible post-translational modification).

    Exposure time : 1 minute

    Image is courtesy of an anonymous AbReview.

    Blocking was with 5% BSA incubated for 1 hour at 25°C.

    See Abreview

  • ab52894 showing positive staining in Glioma tissue.

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

    Lane 1 : H820 siCtl cell lysate, siRNA treated
    Lane 2 : H820 siEGFR#1 cell lysate, siRNA treated
    Lane 3 : H820 siEGFR#2 cell lysate, siRNA treated

    Lysates/proteins at 20 µg per lane.

    Secondary
    GE Healthcare Polyclonal Anti Rabbit, HRP conjugated at 1/5000 dilution

    Predicted band size : 134 kDa
    Observed band size : 165 kDa (why is the actual band size different from the predicted?)

    Gel running conditions: 4-15%, reduced and denatured.

    Blocking agent: 5% milk.

    Blocking time: 1 hour.

    Incubation (with primary antibody): 16 hours, 4 Celsius.

    Dilution buffer: 5%BSA

  • ab52894 showing positive staining in Normal tonsil squamous cells tissue.

  • ab52894 showing positive staining in Renal cell carcinoma tissue.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

Anti-EGFR antibody [EP38Y] (ab52894)参考文献

This product has been referenced in:
  • Chen CC  et al. The matricellular protein CCN1 suppresses hepatocarcinogenesis by inhibiting compensatory proliferation. Oncogene 35:1314-23 (2016). WB . Read more (PubMed: 26028023) »
  • Wang G  et al. Simvastatin induces cell cycle arrest and inhibits proliferation of bladder cancer cells via PPAR? signalling pathway. Sci Rep 6:35783 (2016). WB . Read more (PubMed: 27779188) »

See all 16 Publications for this product

Product Wall

Application Western blot
Sample Human Cell lysate - whole cell (Human Renal Epithelial Cells)
Gel Running Conditions Non-reduced Non-Denaturing (Native) (8%)
Loading amount 60 µg
Treatment 20ng EGF for 30min
Specification Human Renal Epithelial Cells
Blocking step BSA as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Dr. 腾飞 白

Verified customer

提交于 Jan 06 2016

Application Western blot
Sample Mouse Cell lysate - other (macrophage)
Gel Running Conditions Reduced Denaturing
Loading amount 80 µg
Specification macrophage
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Nov 06 2015

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (D3 mouse embryonic stem cells)
Permeabilization Yes - Triton 0.1%
Specification D3 mouse embryonic stem cells
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 1%
Fixative Paraformaldehyde
Username

Miss. Fabiana Luise

Verified customer

提交于 Oct 22 2015

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (4-15%)
Sample Human Cell lysate - whole cell (non small cell lung cancer)
Specification non small cell lung cancer
Treatment siRNA treated
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Oct 07 2014


The pretreatment note in the protocol of the IHC kit refers to the antigen retrieval step that is required for most antibody to achieve a successful IHC-P staining.
For both of these antibodies (ab14440 and ab52894) I can recommend to use Sod...

Read More
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Permeabilization Yes - 0.5% Triton X100 in PBS
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Mar 25 2014

Hello! We do recommend you use an antigen retrieval step, heat mediated such as citrate pH6. The antibody you suggest, ab52894 is a good one. Another EGFR antibody with good feedback is ab9482. We do guarantee our products to work as specified on the d...

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Thank you for contacting us.

Rabbit monoclonals typically have 10-100x the affinity of mouse monoclonals, as such it is often the case that the concentration of a rabbit monoclonal will be lower than a mouse monoclonal. Ab52894 does not ha...

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Paraffin blocks are commonly cut into 4 µm thickness sections and those sections are stored in a sealed box at room temperature for no more than 6 months. We have not observed any particular rapid degradation of the EGFR antigen.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"