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Rabbit Recombinant Monoclonal FAK phospho Y397 antibody. Carrier free. Suitable for WB, ICC/IF and reacts with Human, Mouse, Rat samples. Cited in 7 publications.

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Images

Immunocytochemistry/ Immunofluorescence - Anti-FAK (phospho Y397) antibody [EP2160Y] - Low endotoxin, Azide free (AB223529), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-FAK (phospho Y397) antibody [EP2160Y] - Low endotoxin, Azide free (AB223529), expandable thumbnail
  • Western blot - Anti-FAK (phospho Y397) antibody [EP2160Y] - Low endotoxin, Azide free (AB223529), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBIHC-PICC/IF
Human
Tested
Not recommended
Tested
Mouse
Tested
Not recommended
Expected
Rat
Tested
Not recommended
Expected

Tested
Tested

Species

Human, Mouse, Rat

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

15 products for Alternative Product

2 products for Alternative Version

Target data

Function

Non-receptor protein-tyrosine kinase that plays an essential role in regulating cell migration, adhesion, spreading, reorganization of the actin cytoskeleton, formation and disassembly of focal adhesions and cell protrusions, cell cycle progression, cell proliferation and apoptosis. Required for early embryonic development and placenta development. Required for embryonic angiogenesis, normal cardiomyocyte migration and proliferation, and normal heart development. Regulates axon growth and neuronal cell migration, axon branching and synapse formation; required for normal development of the nervous system. Plays a role in osteogenesis and differentiation of osteoblasts. Functions in integrin signal transduction, but also in signaling downstream of numerous growth factor receptors, G-protein coupled receptors (GPCR), EPHA2, netrin receptors and LDL receptors. Forms multisubunit signaling complexes with SRC and SRC family members upon activation; this leads to the phosphorylation of additional tyrosine residues, creating binding sites for scaffold proteins, effectors and substrates. Regulates numerous signaling pathways. Promotes activation of phosphatidylinositol 3-kinase and the AKT1 signaling cascade. Promotes activation of MAPK1/ERK2, MAPK3/ERK1 and the MAP kinase signaling cascade. Promotes localized and transient activation of guanine nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs), and thereby modulates the activity of Rho family GTPases. Signaling via CAS family members mediates activation of RAC1. Phosphorylates NEDD9 following integrin stimulation (PubMed:9360983). Recruits the ubiquitin ligase MDM2 to P53/TP53 in the nucleus, and thereby regulates P53/TP53 activity, P53/TP53 ubiquitination and proteasomal degradation. Phosphorylates SRC; this increases SRC kinase activity. Phosphorylates ACTN1, ARHGEF7, GRB7, RET and WASL. Promotes phosphorylation of PXN and STAT1; most likely PXN and STAT1 are phosphorylated by a SRC family kinase that is recruited to autophosphorylated PTK2/FAK1, rather than by PTK2/FAK1 itself. Promotes phosphorylation of BCAR1; GIT2 and SHC1; this requires both SRC and PTK2/FAK1. Promotes phosphorylation of BMX and PIK3R1. Isoform 6 (FRNK) does not contain a kinase domain and inhibits PTK2/FAK1 phosphorylation and signaling. Its enhanced expression can attenuate the nuclear accumulation of LPXN and limit its ability to enhance serum response factor (SRF)-dependent gene transcription.Isoform 6Isoform 6 (FRNK) does not contain a kinase domain and inhibits PTK2/FAK1 phosphorylation and signaling. Its enhanced expression can attenuate the nuclear accumulation of LPXN and limit its ability to enhance serum response factor (SRF)-dependent gene transcription.

Alternative names

Recommended products

Rabbit Recombinant Monoclonal FAK phospho Y397 antibody. Carrier free. Suitable for WB, ICC/IF and reacts with Human, Mouse, Rat samples. Cited in 7 publications.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free

Yes

Clone number

EP2160Y

Purification technique

Affinity purification Protein A

Specificity

The activation of phosphorylation of FAK is reported to be related to developmental processes in brain tissue (PMID: 14642275, PMID: 21118706). So expression level of phosphorylated modified FAK in normal brain is quite low causing not easy to be detected. We suggest optimizing experimental protocols (increasing lysate amount, using lower dilution or higher sensitivity ECL substrate) to improve results.

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

Notes

ab223529 is the carrier-free version of Anti-FAK (phospho Y397) antibody [EP2160Y] ab81298.

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

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 batch production

For more information, read more on recombinant antibodies.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

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

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Focal Adhesion Kinase (FAK) also known as Protein Tyrosine Kinase 2 (PTK2) is a non-receptor tyrosine kinase. This protein has a molecular weight of approximately 125 kDa. FAK is expressed at high levels in brain muscle and liver tissues. Mechanically FAK plays a role in cellular adhesion and migration by regulating integrin signaling and cell-extracellular matrix interactions. FAK auto-phosphorylates at tyrosine residue 397 creating a binding site for Src family kinases and promoting downstream signaling pathways.

Biological function summary

Focal Adhesion Kinase participates in the formation of focal adhesions which are complexes that connect the cytoskeleton to the extracellular matrix. The FAK protein functions as an important signaling node in these structures allowing for the assembly of multiprotein signal transduction complexes. FAK also controls cellular processes such as spreading motility and survival. The interaction with proteins such as Src kinases paxillin and talin facilitates its biological roles in cell signaling.

Pathways

Focal Adhesion Kinase engages in the regulation of the MAPK/ERK signaling pathway and the PI3K/AKT pathway. These pathways are instrumental for cell proliferation survival and migration. In these pathways FAK interacts with proteins such as PI3K Grb2 and Sos linking integrin-mediated signals with downstream effects that influence cell behavior and survival.

Associated diseases and disorders

Altered FAK signaling has ties to cancer progression and metastasis as well as cardiovascular diseases. In cancer the overexpression of FAK and its interaction with proteins like Src and VEGFR can drive tumor growth and angiogenesis. In cardiovascular diseases improper FAK activation can lead to aberrant heart tissue remodeling and associated pathologies. Abnormalities in FAK signaling pathways can therefore contribute significantly to the development and progression of these diseases.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

3 product images

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

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

    Unpurified Anti-FAK (phospho Y397) antibody [EP2160Y] ab81298 staining FAK (phospho Y397) in SK-N-SH (human neuroblastoma cell line) cells treated with anandamide (ethanol solution) (Anandamide (ethanol solution), endogenous cannabinoid 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 Anandamide (ethanol solution), endogenous cannabinoid 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 Anti-FAK (phospho Y397) antibody [EP2160Y] 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 (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed 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 (Anti-FAK (phospho Y397) antibody [EP2160Y] ab81298).

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

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

    ICC/IF image of unpurified Anti-FAK (phospho Y397) antibody [EP2160Y] 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 (Anti-FAK (phospho Y397) antibody [EP2160Y] ab81298, 10μg/ml) overnight at +4°C in PBS containing 1% BSA and 0.1% tween. The secondary antibody (green) was Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed 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.

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

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

    Blocking buffer and concentration: 5% NFDM/TBST. 

    Diluting buffer and concentration: 5% NFDM/TBST.

    The activation of phosphorylation of FAK is reported to be related to developmental processes in brain tissue (PMID: 14642275, PMID: 21118706). So expression level of phosphorylated modified FAK in normal brain is quite low causing not easy to be detected. We suggest optimizing experimental protocols (increasing lysate amount, using lower dilution or higher sensitivity ECL substrate) to improve results.

    Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 has been used as a loading control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-FAK (phospho Y397) antibody [EP2160Y] ab81298).

    Lane 1: Untreated NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 20 µg

    Lanes 1 - 5: Western blot - Anti-FAK (phospho Y397) antibody [EP2160Y] (Anti-FAK (phospho Y397) antibody [EP2160Y] ab81298) at 1/1000 dilution

    Lane 2: NIH/3T3 treated with 10mM Pervanadate for 60 min whole cell lysate at 20 µg

    Lane 3: Untreated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 4: HeLa treated with 10mM Pervanadate for 60 min whole cell lysate at 20 µg

    Lane 5: Mouse brain tissue lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 119 kDa

    Observed band size: 119 kDa

    Exposure time: 40s

    Blocking buffer and concentration: 5% NFDM/TBST. 

    Diluting buffer and concentration: 5% NFDM/TBST.

    The activation of phosphorylation of FAK is reported to be related to developmental processes in brain tissue (PMID: 14642275, PMID: 21118706). So expression level of phosphorylated modified FAK in normal brain is quite low causing not easy to be detected. We suggest optimizing experimental protocols (increasing lysate amount, using lower dilution or higher sensitivity ECL substrate) to improve results.

    Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 has been used as a loading control.

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