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Cardiovascular Angiogenesis Adhesion / ECM Integrins
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RecombinantRabMAb

Recombinant Anti-FAK (phospho Y397) antibody [EP2160Y] - Low endotoxin, Azide free (ab223529)

  • Datasheet
  • Certificate of Compliance
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Immunocytochemistry/ Immunofluorescence - Anti-FAK (phospho Y397) antibody [EP2160Y] - Low endotoxin, Azide free (ab223529)
  • Immunocytochemistry/ Immunofluorescence - Anti-FAK (phospho Y397) antibody [EP2160Y] - Low endotoxin, Azide free (ab223529)
  • Immunocytochemistry/ Immunofluorescence - Anti-FAK (phospho Y397) antibody [EP2160Y] - Low endotoxin, Azide free (ab223529)
  • Anti-FAK (phospho Y397) antibody [EP2160Y] - Low endotoxin, Azide free (ab223529)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP2160Y] to FAK (phospho Y397) - Low endotoxin, Azide free
  • Suitable for: WB, ICC/IF
  • Reacts with: Mouse, Rat, Human

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Overview

  • Product name

    Anti-FAK (phospho Y397) antibody [EP2160Y] - Low endotoxin, Azide free
    See all FAK primary antibodies
  • Description

    Rabbit monoclonal [EP2160Y] to FAK (phospho Y397) - Low endotoxin, Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IFmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human FAK.

  • Positive control

    • Human and rat brain tissue lysates SKNSH cell line
  • General notes

    ab223529 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    pH: 7.20
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP2160Y
  • 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

  • Alternative Versions

    • Anti-FAK (phospho Y397) antibody [EP2160Y] - BSA and Azide free (ab271862)
    • Anti-FAK (phospho Y397) antibody [EP2160Y] (ab81298)
  • Compatible Secondaries

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

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant human FAK protein (ab105909)
  • Related Products

    • Anandamide (ethanol solution), endogenous cannabinoid (ab120087)

Applications

Our Abpromise guarantee covers the use of ab223529 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 at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for IHC-P.
  • 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: 5747 Human
      • Entrez Gene: 14083 Mouse
      • Entrez Gene: 25614 Rat
      • Omim: 600758 Human
      • SwissProt: Q05397 Human
      • SwissProt: P34152 Mouse
      • SwissProt: O35346 Rat
      • Unigene: 395482 Human
      • Unigene: 254494 Mouse
      • Unigene: 2809 Rat
      see all
    • 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

    • Immunocytochemistry/ Immunofluorescence - Anti-FAK (phospho Y397) antibody [EP2160Y] - Low endotoxin, Azide free (ab223529)
      Immunocytochemistry/ Immunofluorescence - Anti-FAK (phospho Y397) antibody [EP2160Y] - Low endotoxin, Azide free (ab223529)

      Unpurified ab81298 staining FAK (phospho Y397) in SK-N-SH (human neuroblastoma cell line) cells treated with anandamide (in water soluble emulsion) (ab120429), by ICC/IF. Increase in FAK (phospho Y397) expression correlates with increased concentration of anandamide (in water soluble emulsion), as described in literature.
      The cells were incubated at 37°C for 5 minutes in media containing different concentrations of ab120429 (anandamide (in water soluble emulsion)) in water, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab81298 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. Membranes are stained in red with WGA.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81298).

    • Immunocytochemistry/ Immunofluorescence - Anti-FAK (phospho Y397) antibody [EP2160Y] - Low endotoxin, Azide free (ab223529)
      Immunocytochemistry/ Immunofluorescence - Anti-FAK (phospho Y397) antibody [EP2160Y] - Low endotoxin, Azide free (ab223529)

      Unpurified ab81298 staining FAK (phospho Y397) in SK-N-SH (human neuroblastoma cell line) cells treated with anandamide (ethanol solution) (ab120087), by ICC/IF. Increase in FAK (phospho Y397) expression correlates with increased concentration of anandamide (ethanol solution), as described in literature.
      The cells were incubated at 37°C for 10 minutes in media containing different concentrations of ab120087 (anandamide (ethanol solution)) in ethanol, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab81298 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. Membranes are stained in red with WGA.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81298).

    • Immunocytochemistry/ Immunofluorescence - Anti-FAK (phospho Y397) antibody [EP2160Y] - Low endotoxin, Azide free (ab223529)
      Immunocytochemistry/ Immunofluorescence - Anti-FAK (phospho Y397) antibody [EP2160Y] - Low endotoxin, Azide free (ab223529)

      This ICC/IF data was generated using the same anti-FAK (phospho Y397) antibody clone [EP2160Y] in a different buffer formulation (cat# ab81298).

      ICC/IF image of unpurified ab81298 stained SKNSH cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab81298, 10µg/ml) overnight at +4°C in PBS containing 1% BSA and 0.1% tween. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Anti-FAK (phospho Y397) antibody [EP2160Y] - Low endotoxin, Azide free (ab223529)
      Anti-FAK (phospho Y397) antibody [EP2160Y] - Low endotoxin, Azide free (ab223529)

    Protocols

    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

  • References (7)

    Publishing research using ab223529? Please let us know so that we can cite the reference in this datasheet.

    ab223529 has been referenced in 7 publications.

    • Goodwin JM  et al. An AMPK-independent signaling pathway downstream of the LKB1 tumor suppressor controls Snail1 and metastatic potential. Mol Cell 55:436-50 (2014). PubMed: 25042806
    • Hu Y  et al. Deficiency of Erbin induces resistance of cervical cancer cells to anoikis in a STAT3-dependent manner. Oncogenesis 2:e52 (2013). WB ; Human . PubMed: 23774064
    • You JJ  et al. Cyr61 induces the expression of monocyte chemoattractant protein-1 via the integrin a?ß3, FAK, PI3K/Akt, and NF-?B pathways in retinal vascular endothelial cells. Cell Signal 26:133-140 (2013). PubMed: 24063814
    • Lu H  et al. IGFBP2/FAK pathway is causally associated with dasatinib resistance in non-small cell lung cancer cells. Mol Cancer Ther 12:2864-73 (2013). Human . PubMed: 24130049
    • Xu B  et al. Effect of focal adhesion kinase on the regulation of realignment and tenogenic differentiation of human mesenchymal stem cells by mechanical stretch. Connect Tissue Res 52:373-9 (2011). WB ; Human . PubMed: 21401419
    • Liu L  et al. Extracellular signal-regulated kinase1/2 activated by fluid shear stress promotes osteogenic differentiation of human bone marrow-derived mesenchymal stem cells through novel signaling pathways. Int J Biochem Cell Biol 43:1591-601 (2011). WB ; Human . PubMed: 21810479
    • Barnes EA  et al. Tuberin regulates E-cadherin localization: implications in epithelial-mesenchymal transition. Am J Pathol 177:1765-78 (2010). PubMed: 20813961

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