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Phospho-FAK (Y397) ELISA Kit (ab279795)

Key features and details

  • Sample type: Cell Lysate
  • Detection method: Colorimetric
  • Assay type: Semi-quantitative
  • Reacts with: Human

Overview

  • Product name

    Phospho-FAK (Y397) ELISA Kit
    See all FAK kits

  • Detection method

    Colorimetric

  • Sample type

    Cell Lysate

  • Assay type

    Semi-quantitative

  • Assay duration

    Multiple steps standard assay

  • Species reactivity

    Reacts with: Human

  • Product overview

    Phospho-FAK (Y397) ELISA Kit (ab279795) is a very rapid, convenient, and sensitive assay kit that can monitor the activation or function of important biological pathways in human cell lysates. By determining phosphorylated FAK protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.


    This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-FAK. An anti-pan FAK antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and FAK present in a sample is bound to the wells by the immobilized antibody. The wells are washed and rabbit anti-phospho-FAK (Y397) antibody is used to detect phosphorylated FAK. After washing away unbound antibody, HRP conjugated anti-rabbit IgG is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of FAK (Y397) bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

  • Tested applications

    Suitable for: Sandwich ELISAmore details

  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    20X Wash Buffer Concentrate 1 x 25ml
    2X Cell lysate buffer 1 x 10ml
    5X Assay Diluent 1 x 15ml
    HRP-conjugated anti-rabbit IgG concentrate (500X) 1 vial
    Pan-FAK Coated Microplate 1 unit
    Positive Control - treated Jurkat cell lysate 1 vial
    Rabbit anti-phospho-FAK (Y397)-antibody 2 vials
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml

  • Research areas

  • 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

  • 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

  • Database links

    Images

    • Positive Control
      Positive Control

      Jurkat cells were treated with Pervanadate at 37°C for 10 min.

      Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer.

      Serial dilutions of lysates were analyzed in this ELISA.

    • Jurkat cells treated with/without Pervanadate.
      Jurkat cells treated with/without Pervanadate.
      Jurkat cells were untreated or treated with 1mM Pervanadate for 10 mins.
    • Jurkat cells treated with/without Pervanadate.
      Jurkat cells treated with/without Pervanadate.
      Jurkat cells were untreated or treated with 1mM Pervanadate for 10 mins.

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