Anti-EGFR (phospho Y1068) 抗体 (ab5644)

Reviews (5)Q&A (1)References (38)
Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1068) antibody (ab5644)
  • Western blot - Anti-EGFR (phospho Y1068) antibody (ab5644)
  • Western blot - Anti-EGFR (phospho Y1068) antibody (ab5644)

Key features and details

  • Rabbit polyclonal to EGFR (phospho Y1068)
  • Suitable for: ICC/IF, WB
  • Reacts with: Human
  • Isotype: IgG

リコンビナント抗体で、ロット間での高い再現性を実現

Product image
Anti-EGFR (phospho Y1068) antibody [Y38] (ab32430)
  • 異なるロット間での安定した再現性
  • 容易なスケールアップ
  • 評価試験による特異性の確認済み
  • 倫理基準に準拠 - アニマル・フリーの生産

製品の概要

  • 製品名

    Anti-EGFR (phospho Y1068) antibody
    EGFR 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to EGFR (phospho Y1068)

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: ICC/IF, WBmore details

  • 種交差性

    交差種: Human

  • 免疫原

    Synthetic phosphopeptide (Human) derived from the region of EGFR that contains tyrosine 1068.

  • ポジティブ･コントロール

  • 特記事項

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab5644の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
ICC/IF
1/100.
WB (5)
1/1000. Detects a band of approximately 185 kDa.
特記事項
ICC/IF
1/100.
WB
1/1000. Detects a band of approximately 185 kDa.

ターゲット情報

  • 機能

    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.

  • 組織特異性

    Ubiquitously expressed. Isoform 2 is also expressed in ovarian cancers.

  • 関連疾患

    Lung cancer
    Inflammatory skin and bowel disease, neonatal, 2

  • 配列類似性

    Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.
    Contains 1 protein kinase domain.

  • 翻訳後修飾

    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.

  • 細胞内局在

    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

  • 参照データベース

    • 別名

      • 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

    画像

    • Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1068) antibody (ab5644)
      Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1068) antibody (ab5644)

      Immunofluorescence analysis of A431 (Human epidermoid carcinoma cell line) cells labeling EGFR with ab5644 at 1/100 dilution, followed by Goat anti-rabbit IgG (H+L) Superclonal, Alexa Fluor® 488 conjugate was used as the secondary antibody at 1/2000 dilution (Panel a). Nuclei (Panel b) were stained with SlowFade® Gold Antifade Mountant with DAPI. F-actin (Panel c) was stained with Rhodamine Phalloidin. Panel (d) represents the merged image showing membrane localization. Panel (e) represents cells treated with antagonist, Afatinib (1µM for 6hrs) followed by EGF (200 ng/ml for 10 minutes), showing no Phospho-EGFR staining. Panel (f) shows untreated cells with no signal. Panel (g) represents control cells with no primary antibody to assess background. 

      The cells (in 70% confluent log phase treated with 200ng/ml of EGF for 10 minutes) were fixed with 4% paraformaldehyde for 10 minutes; permeabilized with 0.1% Triton X-100 for 10 minutes and blocked with 1% BSA for hour at room temperature. The images were captured at 60X magnification.

    • Western blot - Anti-EGFR (phospho Y1068) antibody (ab5644)
      Western blot - Anti-EGFR (phospho Y1068) antibody (ab5644)
      All lanes : Anti-EGFR (phospho Y1068) antibody (ab5644) at 1/1000 dilution

      Lane 1 : A431 (Human epidermoid carcinoma cell line) whole cell lysate with skimmed milk
      Lane 2 : A431 whole cell lysate treated with EGF (200 ng/mL for 10 minutes) with skimmed milk
      Lane 3 : A431 whole cell lysate treated with Gefitinib followed by EGF (1uM for 16 hours, 200 ng/mL for 10 minutes) with skimmed milk
      Lane 4 : A431 whole cell lysate treated with Afatinib followed by EGF (0.5 uM for 6 hours, 200 ng/mL for 10 minutes) with skimmed milk
      Lane 5 : A549 (Human lung carcinoma cell line) whole cell lysate with skimmed milk
      Lane 6 : A549 whole cell lysate treated with EGF (200 ng/mL for 10 minutes) with skimmed milk
      Lane 7 : A549 whole cell lysate treated with Afatinib followed by EGF (0.5 uM for 6 hours, 200 ng/mL for 10 minutes) with skimmed milk

      Lysates/proteins at 30 µg per lane.

      Blocking peptides at 5 % per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG Superclonal Secondary Antibody, HRP at 1/4000 dilution


      Western blot analysis using ab5644 shows increased expression of proteins phosphorylated at the tyrosine residues in A-431 and A549 cell lines upon EGF treatment and pre-treatment with EGFR-antagonists, Gefitinib and Afatinib. This results in inhibition of Phospho-EGFR in A-431 and A549 cell lines

    • Western blot - Anti-EGFR (phospho Y1068) antibody (ab5644)
      Western blot - Anti-EGFR (phospho Y1068) antibody (ab5644)This image is courtesy of an Abreview submitted by Mr Samir Nuseibeh
      All lanes : Anti-EGFR (phospho Y1068) antibody (ab5644) at 1/200 dilution

      Lane 1 : A431 (Human epidermoid carcinoma cell line) whole cell lysate - not treated
      Lane 2 : A431 whole cell lysate - 100ng/ml EGF for 1 minute
      Lane 3 : A431 whole cell lysate - 100ng/ml EGF for 2.5 minutes
      Lane 4 : A431 whole cell lysate - 100ng/ml EGF for 5 minutes
      Lane 5 : A431 whole cell lysate - 100ng/ml EGF for 10 minutes
      Lane 6 : A431 whole cell lysate - 100ng/ml EGF for 20 minutes
      Lane 7 : A431 whole cell lysate - 100ng/ml EGF for 40 minutes

      Secondary
      All lanes : HRP conjugated Goat anti-rabbit antibody

      Developed using the ECL technique.

      Performed under reducing conditions.

      Exposure time: 2 minutes

      See Abreview

    参考文献 (38)

    ab5644 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab5644 は 38 報の論文で使用されています。

    • Nakano Y  et al. Clinical importance of long non-coding RNA LINC00460 expression in EGFR-mutant lung adenocarcinoma. Int J Oncol 56:243-257 (2020). PubMed: 31789388
    • Song X  et al. Zi Shen Huo Luo Formula Enhances the Therapeutic Effects of Angiotensin-Converting Enzyme Inhibitors on Hypertensive Left Ventricular Hypertrophy by Interfering With Aldosterone Breakthrough and Affecting Caveolin-1/Mineralocorticoid Receptor Colocalization and Downstream Extracellular Signal-Regulated Kinase Signaling. Front Pharmacol 11:383 (2020). PubMed: 32317965
    • Elkhider A  et al. Aquaporin 5 promotes tumor migration and angiogenesis in non-small cell lung cancer cell line H1299. Oncol Lett 19:1665-1672 (2020). PubMed: 32194658
    • Šuštic T  et al. RUNX2/CBFB modulates the response to MEK inhibitors through activation of receptor tyrosine kinases in KRAS-mutant colorectal cancer. Transl Oncol 13:201-211 (2020). PubMed: 31865182
    • Wang D & Bao B Gallic Acid Impedes Non-Small Cell Lung Cancer Progression via Suppression of EGFR-Dependent CARM1-PELP1 Complex. Drug Des Devel Ther 14:1583-1592 (2020). PubMed: 32425504
    View all Publications for this product

    レビューと Q&A

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    レビューを送る 質問を送る

    1-6 of 6 Abreviews or Q&A

    Western blot abreview for Anti-EGFR (phospho Y1068) antibody

     Below Average
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    50 µg
    Gel Running Conditions
    Reduced Denaturing
    Sample
    Mouse Tissue lysate - whole (heart)
    Specification
    heart
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Ms. sarah shen

    Verified customer

    投稿 May 12 2014

    Western blot abreview for Anti-EGFR (phospho Y1068) antibody

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (Liver)
    Loading amount
    30 µg
    Specification
    Liver
    Gel Running Conditions
    Reduced Denaturing (12)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    投稿 Jun 25 2012

    Western blot abreview for Anti-EGFR (phospho Y1068) antibody

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Tissue lysate - whole (Liver)
    Loading amount
    50 µg
    Specification
    Liver
    Gel Running Conditions
    Reduced Denaturing (12)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    投稿 May 02 2012

    Western blot abreview for Anti-EGFR (phospho Y1068) antibody

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (Liver)
    Loading amount
    50 µg
    Specification
    Liver
    Gel Running Conditions
    Reduced Denaturing (12)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    投稿 Jun 08 2011

    Western blot abreview for Anti-EGFR (phospho Y1068) antibody

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Lung)
    Loading amount
    70 µg
    Specification
    Lung
    Treatment
    EGF 100ng/ml 2.5 minutes
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
    Read More

    Dr. Samir Nuseibeh

    Verified customer

    投稿 Oct 23 2006

    Question

    BATCH NUMBER 154754 ORDER NUMBER unknown DESCRIPTION OF THE PROBLEM Wrong band size. ~75kDa, not 170kDa. Similar results with ab5638 and ab5644. SAMPLE Rat heart tissue lysate. PRIMARY ANTIBODY listed in inquiry DETECTION METHOD Pierce ECL or Pierce Supersignal. POSITIVE AND NEGATIVE CONTROLS USED PC-3 lysate ANTIBODY STORAGE CONDITIONS Aliquotted and stored at -20. SAMPLE PREPARATION Tissue lysis buffer: 50mM HEPES pH7.5 150mM NaCl 1.5mM MgCl2 1mM EGTA 10% glycerol 1% Triton X-100, plus protease inhibitor tablets, plus 10mM NaF and 1mM NaVO4. All done in cold room/on ice. Lysates kept at -80 when not in use. Typical western setup, sample buffer, boil, 7.5% gel. Transferred to PVDF 100V 1 hour, checked transfer with Ponceu S. Blocking/dilution buffer is TBS 2 or 5% BSA fraction V. AMOUNT OF PROTEIN LOADED Usually around 100ug. ELECTROPHORESIS/GEL CONDITIONS reducing & see above TRANSFER AND BLOCKING CONDITIONS see above SECONDARY ANTIBODY Zymed specific to each of the Abcam primaries' species. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 15 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Homogenized with and without phosphatase inhibitors, transfer time, blocking/dilution buffers, incubation temp, incubation volume, antibody concentrations, washings, development time

    Read More
    Answer

    Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. I have been in touch with the originators of ab5638, ab5644 and ab14017 with regards the band sizes that you have detected. I have received the following comments. The mentioned that the most likely explanation for the bands that you have been detecting and the mass of protein that you have been loading is that the the cell preparation that you have been using does not contain any EGFR phosphorylated at these 2 specific sites (ab5638 and ab5644). When the correct target does not exist (or exists in concentrations too low for the antibody to detect), the antibody will bind to either a lower affinity phospho. protein or to the most concentrated protein in the lysate: Neither bindings are specific. The originator also recommended that you try titrating the extracts themselves and the secondary antibody to determine whether the target protein is indeed the protein detected by the antibody or a non-specific band detected by the secondary antiserum. I would like to recommend that you stimulate cells using EGF to induce phosphorylation as shown in the western blot on the datasheet of ab5638 where EGF induced phosphorylation of EGFR. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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