Overview

  • Product nameAnti-FAK (phospho Y397) antibodySee all FAK primary antibodies ...
  • Description
    Rabbit polyclonal to FAK (phospho Y397)
  • Tested applicationsICC/IF, IHC-P, ICC, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Human, Pig, Xenopus laevis, Fruit fly (Drosophila melanogaster), Zebrafish
  • Immunogen

    Synthetic peptide derived from the region of Focal Adhesion Kinase that contains tyrosine 397. The sequence is conserved in human, mouse, rat, chicken and frog.

  • Positive control
    • Chicken embryo fibroblasts expressing Focal Adhesion Kinase protein and plated on fibronectin, and NIH3 T3 cells treated with PDGF.
  • General notes


    Focal Adhesion Kinase is a 125 kDa non-receptor protein tyrosine kinase that was discovered as a substrate for Src, and is a key element of integrin signaling. Focal Adhesion Kinase plays a central role in cell spreading, differentiation, migration, cell death and acceleration of the G1 to S phase transition of the cell cycle. Tyrosine 397 is the autophosphorylation site of Focal Adhesion Kinase. The site binds Src family SH2 domains and the p85 subunit of PI3-Kinase.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage bufferPreservative: 0.05% Sodium Azide
    Constituents: 50% Glycerol, PBS, 1mg/ml BSA (IgG, protease free). pH 7.3
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • Purification notesPurified from rabbit serum by sequential epitope-specific chromatography. The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated Focal Adhesion Kinase. The final product is generated by affinity chromatography using a Focal Adhesion Kinase-derived peptide that is phosphorylated at tyrosine 397.
  • Primary antibody notes Focal Adhesion Kinase is a 125 kDa non-receptor protein tyrosine kinase that was discovered as a substrate for Src, and is a key element of integrin signaling. Focal Adhesion Kinase plays a central role in cell spreading, differentiation, migration, cell death and acceleration of the G1 to S phase transition of the cell cycle. Tyrosine 397 is the autophosphorylation site of Focal Adhesion Kinase. The site binds Src family SH2 domains and the p85 subunit of PI3-Kinase.
  • Clonality Polyclonal
  • IsotypeIgG
  • Research Areas

Applications

Our Abpromise guarantee covers the use of ab4803 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
ICC/IF 1/100.
IHC-P Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
WB 1/1000. Predicted molecular weight: 125 kDa.
IHC-Fr Use at an assay dependent concentration.

Target

  • FunctionNon-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 specificityExpressed in all organs tested, in lymphoid cell lines, but most abundantly in brain.
  • Sequence similaritiesBelongs to the protein kinase superfamily. Tyr protein kinase family. FAK subfamily.
    Contains 1 FERM domain.
    Contains 1 protein kinase domain.
  • DomainThe 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 localizationCell junction > focal adhesion. Cell membrane. Constituent of focal adhesions.
  • Information by UniProt
  • Database links
  • Alternative names
    • FADK 1 antibody
    • FADK antibody
    • FAK 1 antibody
    • FAK 1 antibody
    • FAK related non kinase polypeptide antibody
    • FAK1 antibody
    • FAK1_HUMAN antibody
    • Focal adhesion kinase 1 antibody
    • Focal adhesion Kinase antibody
    • Focal adhesion kinase related nonkinase antibody
    • FRNK antibody
    • p125FAK antibody
    • pp125FAK antibody
    • pp125FAK antibody
    • PPP1R71 antibody
    • Protein phosphatase 1 regulatory subunit 71 antibody
    • Protein tyrosine kinase 2 antibody
    • Protein Tyrosine Kinase Cytoplasmic antibody
    • Protein Tyrosine Kinase Cytoplasmic antibody
    • Protein-tyrosine kinase 2 antibody
    • PTK 2 antibody
    • PTK2 antibody
    • PTK2 protein tyrosine kinase 2 antibody
    • PTK2 protein tyrosine kinase 2 antibody
    see all

Anti-FAK (phospho Y397) antibody images

  • ab4803 staining FAK (phospho Y397) in Zebrafish fin tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at 22°C. Samples were incubated with primary antibody (1/400 in PBS + 0.3% Triton) for 2 hours at 22°C. An undiluted Cy3®-conjugated Donkey anti-rabbit IgG polyclonal was used as the secondary antibody.

    See Abreview



  • Predicted band size : 125 kDa
    Peptide Competition: Extracts prepared from chick embryo fibroblasts expressing FAK and plated on fibronectin were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to nitrocellulose. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4oC, then were incubated with 0.35 µg/mL ab4803 antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1), the non-phosphopeptide corresponding to the immunogen (2), a generic phospho-tyrosine containing peptide (3), a phosphopeptide derived from the corresponding region of Pyk2 (4), or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and bands were detected using the Tropix WesternStar method. The data show that only the peptide corresponding to ab4803 blocks the antibody signal, thereby demonstrating the specificity of the antibody. Peptide Competition: E
  • Detection of ab4803 in focal adhesions of human melanoma cells.
  • ab4803 at 1/100 staining (renin) transgenic rat aorta sections (5 micron) by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). The tissue was paraformaldehyde fixed and blocked with 5% serum prior to incubation with the antibody for 12 hours. A Cy3 ® conjugated goat polyclonal antibody was used as the secondary.

    See Abreview

References for Anti-FAK (phospho Y397) antibody (ab4803)

This product has been referenced in:
  • Shav D  et al. Wall shear stress effects on endothelial-endothelial and endothelial-smooth muscle cell interactions in tissue engineered models of the vascular wall. PLoS One 9:e88304 (2014). ICC/IF ; Human . Read more (PubMed: 24520363) »
  • Kumar B  et al. A novel curcumin analog (H-4073) enhances the therapeutic efficacy of cisplatin treatment in head and neck cancer. PLoS One 9:e93208 (2014). WB ; Human . Read more (PubMed: 24675768) »

See all 24 Publications for this product

Product Wall

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (pancreatic cancer cell lines Panc1, AsPc1)
Specification pancreatic cancer cell lines Panc1, AsPc1
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
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Submitted Jul 25 2014

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Tissue lysate - whole (pancreatic cancer mouse model tissue biosy)
Specification pancreatic cancer mouse model tissue biosy
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
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Submitted Jul 25 2014

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (4-12)
Sample Rat Tissue lysate - whole (Brain)
Specification Brain
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Submitted Jul 24 2014

Application Immunohistochemistry (Frozen sections)
Sample Zebrafish Tissue sections (fin)
Specification fin
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative Paraformaldehyde
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Submitted Nov 18 2013

Application Immunocytochemistry/ Immunofluorescence
Sample Chicken Cell (Oculomotor primary culture)
Specification Oculomotor primary culture
Fixative Paraformaldehyde
Permeabilization No
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
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Submitted Feb 07 2013

Thank you for contacting us.

We do offer theFAK peptide - phospho Y397. The product number is ab40145 and may be located at the following link:

http://www.abcam.com/FAK-peptide-phospho-Y397-ab40145.html

I hope this ...

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Currently, we do not have any in-house data regarding the specificity of this antibody for particular isoforms of FAK . I would expect that this antibody would recognize any isoform of human, rat ,mouse, chicken or frog FAK that is phosphoryl...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (IEC6)
Loading amount 25 µg
Specification IEC6
Gel Running Conditions Reduced Non-Denaturing (Native) (4-20%)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 24°C
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Submitted Sep 10 2012

According to our records, ab76496 and ab4803was proving difficult to use inWB and we were in contact in order to help resolve the issue.

Looking at our correspondence, it appears that we are awaiting more details in order to help us better und...

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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...

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