製品の概要

  • 製品名
    Anti-FAK (phospho Y397) antibody
    FAK 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to FAK (phospho Y397)
  • 特異性
    This antibody gave a positive result in ELISA against the immunizing peptide (ab40145). It gave a negative result in ELISA against the non-modified equivalent peptide (ab53601). This indicates that it is specific for the modified peptide.
  • アプリケーション
    適用あり: WB, ICC/IF, IHC-Pmore details
  • 種交差性
    交差種: Mouse, Human, Drosophila melanogaster
    交差が予測される動物種: Rat, Chicken, Zebrafish
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 350 - 450 of Human FAK, phosphorylated at Y397.

    (Peptide available as ab40145.)

  • ポジティブ・コントロール
    • NIH 3T3 Whole Cell Lysate, NIH 3T3 Whole Cell Lysate treated with Vanadate + PDGF, A431 Whole Cell Lysate trated with EGF

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab39967 in the following tested applications.

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

アプリケーション Abreviews 特記事項
WB Use a concentration of 1 µg/ml. Detects a band of approximately 119 kDa (predicted molecular weight: 119 kDa).
ICC/IF Use a concentration of 5 µg/ml.
IHC-P Use at an assay dependent concentration.

ターゲット情報

  • 機能
    Non-receptor protein-tyrosine kinase implicated in signaling pathways involved in cell motility, proliferation and apoptosis. Activated by tyrosine-phosphorylation in response to either integrin clustering induced by cell adhesion or antibody cross-linking, or via G-protein coupled receptor (GPCR) occupancy by ligands such as bombesin or lysophosphatidic acid, or via LDL receptor occupancy. Microtubule-induced dephosphorylation at Tyr-397 is crucial for the induction of focal adhesion disassembly. Plays a potential role in oncogenic transformations resulting in increased kinase activity.
  • 組織特異性
    Expressed in all organs tested, in lymphoid cell lines, but most abundantly in brain.
  • 配列類似性
    Belongs to the protein kinase superfamily. Tyr protein kinase family. FAK subfamily.
    Contains 1 FERM domain.
    Contains 1 protein kinase domain.
  • ドメイン
    The first Pro-rich domain interacts with the SH3 domain of CRK-associated substrate (BCAR1) and CASL.
    The carboxy-terminal region is the site of focal adhesion targeting (FAT) sequence which mediates the localization of FAK1 to focal adhesions.
  • 翻訳後修飾
    Phosphorylated on 6 tyrosine residues upon activation. Microtubule-induced dephosphorylation at Tyr-397 could be catalyzed by PTPN11 and regulated by ZFYVE21. Dephosphorylated by PTPN11 upon EPHA2 activation by its ligand EFNA1.
  • 細胞内局在
    Cell junction > focal adhesion. Cell membrane. Constituent of focal adhesions.
  • Information by UniProt
  • 参照データベース
  • 別名
    • FADK 1 antibody
    • FADK antibody
    • FAK related non kinase polypeptide antibody
    • FAK1 antibody
    • FAK1_HUMAN antibody
    • Focal adhesion kinase 1 antibody
    • Focal adhesion Kinase antibody
    • Focal adhesion kinase isoform FAK Del33 antibody
    • Focal adhesion kinase related nonkinase antibody
    • FRNK antibody
    • p125FAK antibody
    • pp125FAK antibody
    • PPP1R71 antibody
    • Protein phosphatase 1 regulatory subunit 71 antibody
    • Protein tyrosine kinase 2 antibody
    • Protein-tyrosine kinase 2 antibody
    • Ptk2 antibody
    • PTK2 protein tyrosine kinase 2 antibody
    see all

画像

  • All lanes : Anti-FAK (phospho Y397) antibody (ab39967) at 1 µg/ml

    Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg
    Lane 2 : NIH 3T3 treated with Vanadate and PDGF Whole Cell Lysate at 10 µg
    Lane 3 : EGF-Stimulated A431 Whole Cell Lysate at 20 µg
    Lane 4 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg with Mouse FAK (phospho Y397) peptide (ab40145) at 1 µg/ml
    Lane 5 : NIH 3T3 treated with Vanadate and PDGF Whole Cell Lysate at 10 µg with Mouse FAK (phospho Y397) peptide (ab40145) at 1 µg/ml
    Lane 6 : EGF-Stimulated A431 Whole Cell Lysate at 20 µg with Mouse FAK (phospho Y397) peptide (ab40145) at 1 µg/ml
    Lane 7 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg with Mouse FAK peptide (ab53601) at 1 µg/ml
    Lane 8 : NIH 3T3 treated with Vanadate and PDGF Whole Cell Lysate at 10 µg with Mouse FAK peptide (ab53601) at 1 µg/ml
    Lane 9 : EGF-Stimulated A431 Whole Cell Lysate at 20 µg with Mouse FAK peptide (ab53601) at 1 µg/ml

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 119 kDa
    Observed band size : 120 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 115 kDa,200 kDa,90 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 4 minutes

    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 using 2% Bovine Serum Albumin before being incubated with ab39967 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • ICC/IF image of ab39967 stained human HeLa cells. The cells were methanol fixed (5 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab39967, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
  • All lanes : Anti-FAK (phospho Y397) antibody (ab39967) at 1 µg/ml

    Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 2 : NIH 3T3 treated with Vanadate and PDGF Whole Cell Lysate
    Lane 3 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate with Mouse FAK (phospho Y397) peptide (ab40145) at 1 µg/ml
    Lane 4 : NIH 3T3 treated with Vanadate and PDGF Whole Cell Lysate with Mouse FAK (phospho Y397) peptide (ab40145) at 1 µg/ml
    Lane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate with Mouse FAK peptide (ab53601) at 1 µg/ml
    Lane 6 : NIH 3T3 treated with Vanadate and PDGF Whole Cell Lysate with Mouse FAK peptide (ab53601) at 1 µg/ml

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 119 kDa
    Observed band size : 119 kDa

参考文献

This product has been referenced in:
  • McKay TB  et al. Acute hypoxia influences collagen and matrix metalloproteinase expression by human keratoconus cells in vitro. PLoS One 12:e0176017 (2017). WB ; Human . Read more (PubMed: 28426715) »
  • Sero JE & Bakal C Multiparametric Analysis of Cell Shape Demonstrates that ß-PIX Directly Couples YAP Activation to Extracellular Matrix Adhesion. Cell Syst 4:84-96.e6 (2017). Read more (PubMed: 28065575) »

See all 8 Publications for this product

レビューと Q&A

Filter by Application

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Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 15 minute(s) · Concentration: 1% · Temperature: 21°C
Sample
Pig Cell (Porcine aortic endothelial cell)
Specification
Porcine aortic endothelial cell
Permeabilization
Yes - 0.1% Triton X-100
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Mar 23 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Mouse Embryonic Fibroblast)
Permeabilization
Yes - 0.5% TX-100
Specification
Mouse Embryonic Fibroblast
Blocking step
Serum as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Fixative
Paraformaldehyde
Username

Samantha O'Hara

Verified customer

投稿 Nov 18 2015

Application
Western blot
Loading amount
4.7 µg
Gel Running Conditions
Reduced Denaturing (12% Bis-Tris gel)
Sample
Pig Cell lysate - whole cell (Porcine primary aortic endothelial cells)
Specification
Porcine primary aortic endothelial cells
Treatment
30 ng/mL genistein for 0, 1, or 2 hours
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 21°C
Username

Abcam user community

Verified customer

投稿 Mar 16 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (spinal chord)
Specification
spinal chord
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Dako Target Retreival Solution
Permeabilization
No
Blocking step
5 minutes of peroxidase block then 10 minutes of protein block. These are ready-to-use reagents purchased from Dako as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 20°C
Username

Mr. Antibody Solutions

Verified customer

投稿 Jun 01 2010

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

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