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  1. Link

    mouse-fak-phospho-y397-peptide-ab40145.pdf

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Cardiovascular Angiogenesis Adhesion / ECM Integrins
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Mouse FAK (phospho Y397) peptide (ab40145)

  • Datasheet
Reviews (1)Q&A (1)References (3)

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Blocking - Mouse FAK (phospho Y397) peptide (ab40145)
  • Western blot - Mouse FAK (phospho Y397) peptide (ab40145)

Key features and details

  • Purity: > 90% HPLC
  • Suitable for: Blocking

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Anti-FAK (phospho Y397) antibody [EP2160Y] (ab81298)

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Description

  • Product name

    Mouse FAK (phospho Y397) peptide
    See all FAK proteins and peptides
  • Purity

    > 90 % HPLC.

  • Accession

    P34152
  • Animal free

    No
  • Nature

    Synthetic
    • Species

      Mouse
    • Modifications

      phospho Y397

Specifications

Our Abpromise guarantee covers the use of ab40145 in the following tested applications.

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

  • Applications

    Blocking

  • Form

    Liquid
  • Additional notes

    - First try to dissolve a small amount of peptide in either water or buffer. The more charged residues on a peptide, the more soluble it is in aqueous solutions.
    - If the peptide doesn’t dissolve try an organic solvent e.g. DMSO, then dilute using water or buffer.
    - Consider that any solvent used must be compatible with your assay. If a peptide does not dissolve and you need to recover it, lyophilise to remove the solvent.
    - Gentle warming and sonication can effectively aid peptide solubilisation. If the solution is cloudy or has gelled the peptide may be in suspension rather than solubilised.
    - Peptides containing cysteine are easily oxidised, so should be prepared in solution just prior to use.

  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

    Information available upon request.

General Info

  • Alternative names

    • FADK
    • FADK 1
    • FAK related non kinase polypeptide
    • FAK1
    • FAK1_HUMAN
    • Focal adhesion Kinase
    • Focal adhesion kinase 1
    • Focal adhesion kinase isoform FAK Del33
    • Focal adhesion kinase related nonkinase
    • FRNK
    • p125FAK
    • pp125FAK
    • PPP1R71
    • Protein phosphatase 1 regulatory subunit 71
    • Protein tyrosine kinase 2
    • Protein-tyrosine kinase 2
    • Ptk2
    • PTK2 protein tyrosine kinase 2
    see all
  • 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.
  • Target information above from: UniProt accession Q05397 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt

Images

  • Blocking - Mouse FAK (phospho Y397) peptide (ab40145)
    Blocking - Mouse FAK (phospho Y397) peptide (ab40145)This image is courtesy of an anonymous Abreview

    To demonstrate the specificity of the phosphorylation site specific antibody (ab4803) a 1/150 dilution of anti human-FAK (phospho Y397) antibody was pre-incubated with 200 fold molar excess of peptide (ab40145, 1/15 dilution) for 12 hours rolling at 4°C.

    Human Tissue sections (kidney) were deparaffinized, rehydrated and antigen retrieval was performed using citrate buffer pH 6.0. Endogenous peroxidase was quenched using 3% hydrogenperoxide in methanol and blocking was performed. Primary antibody alone (positive control) or mixed with peptide (blocking control) or antibody diluent alone (negative control) was applied to tissue sections and incubated for 16 hours at 4°C. Detection was performed using a HRP detection system. Nuclei were counterstained using Hematoxylin.

  • Western blot - Mouse FAK (phospho Y397) peptide (ab40145)
    Western blot - Mouse FAK (phospho Y397) peptide (ab40145)
    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
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Observed band size: 119 kDa why is the actual band size different from the predicted?

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

References (3)

Publishing research using ab40145? Please let us know so that we can cite the reference in this datasheet.

ab40145 has been referenced in 3 publications.

  • Zaghdoudi S  et al. FAK activity in cancer-associated fibroblasts is a prognostic marker and a druggable key metastatic player in pancreatic cancer. EMBO Mol Med 12:e12010 (2020). PubMed: 33025708
  • Diaz Osterman CJ  et al. FAK activity sustains intrinsic and acquired ovarian cancer resistance to platinum chemotherapy. Elife 8:N/A (2019). PubMed: 31478830
  • Aust S  et al. Ambivalent role of pFAK-Y397 in serous ovarian cancer--a study of the OVCAD consortium. Mol Cancer 13:67 (2014). PubMed: 24655477

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Peptide Competition Experiment

Excellent
Abreviews
Abreviews
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Application
Immunohistochemistry
To demonstrate the specificity of the phosphorylation site specific antibody (ab4803) a 1/150 dilution of anti human-FAK (phospho Y397) antibody in DAKO AB diluent was pre-incubated with 200 fold molar excess of peptide (1/15 dilution) for 12h rolling at 4°C.
Human Tissue sections (kidney) were deparaffinized, rehydrated and antigen retrieval was performed using citrate buffer pH 6.0. Endogenous peroxidase was quenched using 3% hydrogenperoxide in methanol and blocking was performed with ultra V block (Thermo Scientific™). Primary antibody alone (positive control) or mixed with peptide (blocking control) or antibody diluent alone (negative control) was applied to tissue sections and incubated for 16h at 4°C. Detection was performed using Ultravision LP Detection System (Thermo Scientific™) with HRP. Nuclei were counterstained using Hematoxylin.

Abcam user community

Verified customer

Submitted Mar 12 2014

Question

Is there a peptide for use with this antibody as a positive control.

Read More

Abcam community

Verified customer

Asked on Oct 05 2012

Answer

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:

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

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

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Abcam Scientific Support

Answered on Oct 05 2012

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