Overview

  • Product name
    Anti-FAK (phospho Y397) antibody
    See all FAK primary antibodies
  • Description
    Rabbit polyclonal to FAK (phospho Y397)
  • Specificity
    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.
  • Tested applications
    Suitable for: WB, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human, Drosophila melanogaster
    Predicted to work with: Rat, Chicken, Zebrafish
  • Immunogen

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

    (Peptide available as ab40145.)

  • Positive control
    • NIH 3T3 Whole Cell Lysate, NIH 3T3 Whole Cell Lysate treated with Vanadate + PDGF, A431 Whole Cell Lysate trated with EGF

Properties

Applications

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.

Application Abreviews Notes
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.

Target

  • Function
    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.
  • Tissue specificity
    Expressed in all organs tested, in lymphoid cell lines, but most abundantly in brain.
  • Sequence similarities
    Belongs to the protein kinase superfamily. Tyr protein kinase family. FAK subfamily.
    Contains 1 FERM domain.
    Contains 1 protein kinase domain.
  • Domain
    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.
  • Post-translational
    modifications
    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.
  • Cellular localization
    Cell junction > focal adhesion. Cell membrane. Constituent of focal adhesions.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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

Anti-FAK (phospho Y397) antibody images

  • 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

References for Anti-FAK (phospho Y397) antibody (ab39967)

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

Product Wall

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

Submitted 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

Submitted 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

Submitted Mar 16 2015

Thank you for contacting us.

All of these products are tested in IHC-P and are fully guarnateed. Anyone from these can be chosen.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more adv...

Read More
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

Submitted Jun 01 2010

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