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Cardiovascular Angiogenesis Adhesion / ECM Integrins
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Anti-FAK (phospho Y861) antibody (ab4804)

  • Datasheet
Reviews (2)Q&A (1)References (3)

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Western blot - Anti-FAK (phospho Y861) antibody (ab4804)
  • Western blot - Anti-FAK (phospho Y861) antibody (ab4804)

Key features and details

  • Rabbit polyclonal to FAK (phospho Y861)
  • Suitable for: WB
  • Reacts with: Chicken
  • Isotype: IgG

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Protein
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Recombinant human FAK protein (ab105909)
Secondary
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Knockout
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Human PTK2 (FAK) knockout HEK-293T cell line (ab255421)

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Overview

  • Product name

    Anti-FAK (phospho Y861) antibody
    See all FAK primary antibodies
  • Description

    Rabbit polyclonal to FAK (phospho Y861)
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    WB
    Chicken
    See all applications and species data
  • Immunogen

    Synthetic peptide (Human) derived from the region of human Focal Adhesion Kinase that contains tyrosine 861. The sequence is conserved in human, mouse, rat, chicken and frog.

  • General notes


    Focal Adhesion Kinase is a 125 kDa non-receptor protein tyrosine kinase that is a substrate for Src and a key element in growth factor and integrin signalling. 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. Tyr861 of Focal Adhesion Kinase is a major Src phosphorylation site that allows Focal Adhesion Kinase to bind to integrins and is also involved in cancer.

Properties

  • Form

    Liquid
  • Storage instructions

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

    pH: 7.3
    Preservative: 0.05% Sodium azide
    Constituents: PBS, 50% Glycerol (glycerin, glycerine), 0.1% BSA

    BSA is IgG and protease free
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    Purified from rabbit serum by sequential epitope-specific chromatography. The antibody has been negatively preadsorbed using (i) a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated Focal Adhesion Kinase protein, and (ii) a generic tyrosine phosphorylated peptide to remove antibody that is reactive with phosphotyrosine (irrespective of the sequence). The final product is generated by affinity chromatography using a Focal Adhesion Kinase-derived peptide that is phosphorylated at tyrosine 861.
  • Primary antibody notes

    Focal Adhesion Kinase is a 125 kDa non-receptor protein tyrosine kinase that is a substrate for Src and a key element in growth factor and integrin signalling. 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. Tyr861 of Focal Adhesion Kinase is a major Src phosphorylation site that allows Focal Adhesion Kinase to bind to integrins and is also involved in cancer.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Angiogenesis
    • Adhesion / ECM
    • Integrins
    • Signal Transduction
    • Protein Phosphorylation
    • Tyrosine Kinases
    • FAK / PYK
    • Signal Transduction
    • Cytoskeleton / ECM
    • Extracellular Matrix
    • Structures
    • Focal Adhesions
    • Cancer
    • Signal transduction
    • Protein phosphorylation
    • Tyrosine kinases
    • FAK family

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant human FAK protein (ab105909)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab4804 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
WB
Chicken
All applications
Rat
Xenopus laevis
Application Abreviews Notes
WB (2)
Use at an assay dependent concentration. Predicted molecular weight: 119 kDa.
Notes
WB
Use at an assay dependent concentration. Predicted molecular weight: 119 kDa.

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.
  • 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
  • Database links

    • Entrez Gene: 396416 Chicken
    • Entrez Gene: 25614 Rat
    • Entrez Gene: 399286 Xenopus laevis
    • SwissProt: Q00944 Chicken
    • SwissProt: O35346 Rat
    • SwissProt: Q91738 Xenopus laevis
    • Unigene: 2809 Rat
    • Unigene: 6819 Xenopus laevis
    • 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

    • Western blot - Anti-FAK (phospho Y861) antibody (ab4804)
      Western blot - Anti-FAK (phospho Y861) antibody (ab4804)
      Peptide Competition: Cell extracts prepared from chick embryo fibroblasts expressing FAK and plated on fibronectin were resolved by SDS-PAGE on a 10% Tris-glycine gel. The proteins then were transferred to nitrocellulose and incubated with 0.50 µg/mL ab4804 antibody, following prior incubation with: (1) the phosphopeptide immunogen, (2) a generic phosphotyrosine containing peptide, (3) the non-phosphorylated peptide corresponding to the phosphopeptide, and (4) no peptide. After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and bands were detected using the Tropix WesternStar detection method. The data show that only the phosphopeptide corresponding to this site blocks the antibody signal, demonstrating the specificity of the ab4804 antibody for this phosphorylated residue. Peptide Competition: Cell extracts prepared from chick embryo fibroblasts expressing FAK and plated on fibronectin were resolved by SDS-PAGE on a 10% T
    • Western blot - Anti-FAK (phospho Y861) antibody (ab4804)
      Western blot - Anti-FAK (phospho Y861) antibody (ab4804)Image courtesy of an anonymous Abreview.
      All lanes : Anti-FAK (phospho Y861) antibody (ab4804) at 1/1000 dilution

      Lane 1 : Whole cell lysate prepared from human MDA-MB-231 breast cancer cells, un-treated
      Lane 2 : Whole cell lysate prepared from human MDA-MB-231 breast cancer cells, treated for 1hr with 2.5uM AZD0530 src inhibitor
      Lane 3 : Whole cell lysate prepared from human MDA-MB-231 breast cancer cells, treated for 1hr with 5uM AZD0530 src inhibitor

      Lysates/proteins at 25 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit HRP conjugated at 1/5000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 119 kDa
      Observed band size: 125 kDa
      why is the actual band size different from the predicted?


      Exposure time: 5 minutes


      Primary antibody incubated for 16 hours at 4°C.
      Blocking step was performed using 5% milk for 1 hour at 25°C.

      See Abreview

    Protocols

    • Peptide Competition Experiment

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
  • References (3)

    Publishing research using ab4804? Please let us know so that we can cite the reference in this datasheet.

    ab4804 has been referenced in 3 publications.

    • 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). PubMed: 25471072
    • Ding LW  et al. LNK (SH2B3): paradoxical effects in ovarian cancer. Oncogene N/A:N/A (2014). PubMed: 24704825
    • Tan CL  et al. Integrin activation promotes axon growth on inhibitory chondroitin sulfate proteoglycans by enhancing integrin signaling. J Neurosci 31:6289-95 (2011). ICC/IF ; Human . PubMed: 21525268

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-3 of 3 Abreviews or Q&A

    Western blot abreview for Anti-FAK (phospho Y861) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Human Synoviocyte fibroblast-like cell)
    Gel Running Conditions
    Reduced Denaturing (8% SDS-PAGE)
    Loading amount
    20 µg
    Treatment
    High glucose 500mg/dl D-glucose for 24hrs
    Specification
    Human Synoviocyte fibroblast-like cell
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 27 2016

    Question

    Wir hatten von Abcam vor einiger Zeit einen Antikörper bestellt, um diesen
    in Maus zu testen da er für humanes ApoER2 ausgeschrieben ist.
    Dies hatten wir nun auch getan, allerdings ohne Erfolg (siehe Anhang).
    Nun wollten wir dieses Ergebnis auf der Abcam Homepage hoch laden,
    allerdings bin ich mir nicht sicher ob ich bei der richtigen Seite gelandet
    bin. Ich konnte mir einen Account anlegen, allerdings habe ich nichts
    gefunden wo ich den DISCOUNT CODE eingeben konnte, und ich weiß nicht wo es
    genau hervor geht, dass dieser Antikörper zum Testen bestellt wurde.
    Können Sie mir hierbei weiterhelfen?
    Wir haben das Gefühl dass der Antikörper generell nicht funktioniert, da die
    humane und murine ApoER2 Ektodomäne eine Sequenzhomologie von über 80%
    aufweisen. Aus diesem Grund ist es komisch, dass so gar kein Signal zu
    detektieren war. Allerdings haben wir für den Abcam ApoER2 Antikörper keine
    positiv Kontrolle, da wir kein humanes ApoER2 Konstrukt haben.
    Leider ist es uns in letzter Zeit mit einem weiteren Abcam Antikörper so
    ergangen, dass er nicht funktionierte (pFAK; ab4804). Dieser weist zwar eine
    Bande auf aber nicht bei den vorhergesagten 125 kDa, sondern bei einer Höhe
    von ca. 95 kDa.

    Mit freundlichen Grüßen

    Read More

    Abcam community

    Verified customer

    Asked on Oct 19 2012

    Answer

    Vielen Dank für Ihre Anfrage und Ihr Feedback.

    Um den Gutschein Code einlösen zu können, müssen Sie eine Abreview verfassen. Auf dem Datenblatt von ab58216 gibt es mehrere Links über die das gemacht werden kann:

    Direkt unter dem Antikörpernamen oder wenn Sie auf den reiter (tab) "Abreviews" klicken und dann "submit an Abreview".

    Bitte geben Sie den Gutschein Code in den "additional notes" in der Abreviewform an damit er aktiviert wird, wenn der Abreview eingereicht wird.

    Danach kann der Code auf einer neuen Bestellung benutzt. werden.

    Es tut mir leid zu hören, dass Sie Probleme mit ab4804 in WB haben.

    Ich habe unseren Fragebogen als Word-Dokument an diese E-Mail angehängt. Durch das Ausfüllen des Fragebogens erhalten wir alle nötigen Informationen über Ihre Proben und Ihr Protokoll. Sobald Sie dieses Formular an uns zurückgeschickt haben, werden wir uns Ihr Protokoll ansehen und möglichst Veränderungsvorschläge machen, die Ihre Ergebnisse verbessern werden.

    Für diesen Antikörper gibt es ein speziell empfohlenes Protokoll:

    https://www.abcam.com/assets/popups/popup_datasheet_protocols_printer_friendly.cfm?pid=310&intAbId=4804

    Falls sich herausstellt, dass der Antikörper nicht so funktioniert, wie auf dem Datenblatt beschrieben und er innerhalb der letzten sechs Monaten gekauft wurde, werden wir Ihnen gerne einen Ersatz oder eine Gutschrift schicken.
    Ich freue mich, bald wieder von Ihnen zu hören.

    Read More

    Abcam Scientific Support

    Answered on Oct 19 2012

    Western blot abreview for Anti-FAK (phospho Y861) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (MDA-MB-231 human breast cancer cells)
    Loading amount
    25 µg
    Specification
    MDA-MB-231 human breast cancer cells
    Treatment
    either untreated or treated with 2.5 or 5uM src inhibitor AZD0530
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 17 2010

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