Overview

  • Product name
    Anti-FAK (phospho Y925) antibody
    See all FAK primary antibodies
  • Description
    Rabbit polyclonal to FAK (phospho Y925)
  • Host species
    Rabbit
  • Specificity
    This antibody is specific for FAK only when phosphorylated at tyrosine 925.
  • Tested applications
    Suitable for: WB, ELISA, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic phosphopeptide derived from Human FAK around the phosphorylation site of tyrosine 925 (KVYpEN).

  • Positive control
    • HepG2 and 293 cell extract.

Properties

Applications

Our Abpromise guarantee covers the use of ab38512 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 1/500 - 1/1000. Detects a band of approximately 123 kDa (predicted molecular weight: 119 kDa).
ELISA 1/4000.
ICC/IF Use at an assay dependent concentration. PubMed: 22102809

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

Images

  • All lanes : Anti-FAK (phospho Y925) antibody (ab38512) at 1/500 dilution

    Lane 1 : HepG2 cell lysate, pre-incubated with phosphopeptide.
    Lane 2 : HepG2 cell lysate.
    Lane 3 : 293 cell lysate + EGF + serum.

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : Alkaline Phosphatase AffiniPure Goat Anti-Rabbit IgG (H+L).

    Predicted band size: 119 kDa
    Observed band size: 123 kDa
    why is the actual band size different from the predicted?



    Lanes can be loaded with 5-30µg of total protein.
  • ab38512 staining FAK (phospho Y925) in murine bone marrow-derived dendritic cells by Immunocytochemistry/ Immunofluorescence.

    Cells were fixed with formaldehyde and blocked with 3% BSA before being incubated with primary antibody. An AlexaFluor®568-conjugated goat anti-rabbit polyclonal IgG was used as the secondary antibody.

References

This product has been referenced in:
  • Cui S  et al. Genistein inhibits the growth and regulates the migration and invasion abilities of melanoma cells via the FAK/paxillin and MAPK pathways. Oncotarget 8:21674-21691 (2017). Read more (PubMed: 28423510) »
  • Gill MB  et al. KSHV-TK is a tyrosine kinase that disrupts focal adhesions and induces Rho-mediated cell contraction. EMBO J 34:448-65 (2015). Read more (PubMed: 25471072) »
See all 6 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer

I am still waiting for the alignment result for ab131349, but our source for ab38512 tells us that it is "designed to react with human FAK phosphorylated at Y925, mouse FAK phosphorylated at Y963, and rat FAK phosphorylated at Y928. It does not react with pig FAK phosphorylated at Y925 according to the alignment of the immunogen with the predicted protein sequence, (http://www.ncbi.nlm.nih.gov/protein/XP_001926033.2)."

It appears that there is no tyrosine at or near amino acid position 925 in the predicted sequence, so the above statement is a little mis-leading but in any case, it is unlikely ab38512 will react with pig phospho-FAK. I expect that will be the case for ab131349 too, but I will confirm, hopefully by early next week at the latest.

Read More

Answer

Thank you for your enquiry. I am sorry to hear you are having a problem with ab38512 (FAK (phospho Y925) antibody). Along with some questions, I would like to suggest the following modifications to your protocol to help increase the detection of FAK - phospho Y925: 1) How much ab38512 was used and how was the incubation performed? The recommended dilution for WB is ~ 1:500 and I suggest incubating the membrane at 4C overnight. 2) Include a recommended positive control, HepG2 or 293. 3) Can you clarify "SKOV3 cells itself + ve" in the controls used? 4) Lyse cells using Tris-Triton or RIPA buffer + fresh cocktail of protease inhibitors - 15 min spinning at 4C. Did you use orthrovanadate or other phosphatase inhibitors in preparing your protein samples? 5) How long was the membrane blocked for? Have you tried other blocking reagents, BSA or serum? Try to block for 2hrs at room temperature. 6) How was the washing steps performed? I recommend 4X 5 min rocking in TPBS. Please let me know if this improves your results. I look forward to your reply.

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