Recombinant Anti-FAK (phospho Y397) antibody [EP2160Y] (ab81298)
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)
- Suitable for: WB, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-FAK (phospho Y397) antibody [EP2160Y]
See all FAK primary antibodies -
Description
Rabbit monoclonal [EP2160Y] to FAK (phospho Y397) -
Host species
Rabbit -
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.
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Tested applications
Suitable for: WB, ICC/IFmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human, mouse and rat brain tissue, treated NIH/3T3 with 10mM Pervanadate for 60 min and treated HeLa with 10mM Pervanadate for 60 min cell lysates. ICC/IF: SK-N-SH cell line.
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General notes
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP2160Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab81298 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 |
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WB | (1) |
1/1000. Predicted molecular weight: 119 kDa.
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ICC/IF | (1) |
Use a concentration of 5 µg/ml.
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Notes |
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WB
1/1000. Predicted molecular weight: 119 kDa. |
ICC/IF
Use a concentration of 5 µg/ml. |
Target
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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
modificationsPhosphorylated 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
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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
see all -
Alternative names
- FADK 1 antibody
- FADK antibody
- FAK related non kinase polypeptide antibody
see all
Images
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All lanes : Anti-FAK (phospho Y397) antibody [EP2160Y] (ab81298) at 1/1000 dilution
Lane 1 : Untreated NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lane 2 : NIH/3T3 treated with 10mM Pervanadate for 60 min whole cell lysate
Lane 3 : Untreated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 4 : HeLa treated with 10mM Pervanadate for 60 min whole cell lysate
Lane 5 : Mouse brain tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 119 kDa
Observed band size: 119 kDa
Exposure time: 40 secondsBlocking 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.
ab181602 has been used as a loading control.
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All lanes : Anti-FAK (phospho Y397) antibody [EP2160Y] (ab81298) at 1/2000 dilution (purified)
Lane 1 : Untreated mouse brain
Lane 2 : Mouse brain treated with alkaline phosphatase
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 119 kDa
Observed band size: 119 kDaBlocking Buffer: 5% NFDM/TBST
Dilution Buffer: 5% NFDM/TBST
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ICC/IF image of unpurified ab81298 stained SK-N-SH (human neuroblastoma cell line) 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.
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All lanes : Anti-FAK (phospho Y397) antibody [EP2160Y] (ab81298) at 1/1000 dilution (purified)
Lane 1 : Untreated rat brain
Lane 2 : Rat brain treated with alkaline phosphatase
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP conjugated goat anti-rabbit IgG (H+L) at 1000 µg
Predicted band size: 119 kDa
Observed band size: 119 kDaBlocking Buffer: 5% NFDM/TBST
Dilution Buffer: 5% NFDM/TBST
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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. -
All lanes : Anti-FAK (phospho Y397) antibody [EP2160Y] (ab81298) at 1/2000 dilution (unpurified)
Lane 1 : human brain tissue lysates, untreated.
Lane 2 : human brain tissue lysates treated with AP.
Lane 3 : rat brain tissue lysates, untreated.
Lane 4 : rat brain tissue lysates treated with AP.
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti rabbit. at 1/2000 dilution
Predicted band size: 119 kDa
Observed band size: 119 kDa -
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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (118)
ab81298 has been referenced in 118 publications.
- Yu L et al. Carboxymethyl chitosan-alginate enhances bone repair effects of magnesium phosphate bone cement by activating the FAK-Wnt pathway. Bioact Mater 20:598-609 (2023). PubMed: 35846837
- Pang S et al. Integrin β1/FAK/SRC signal pathway is involved in autism spectrum disorder in Tspan7 knockout rats. Life Sci Alliance 6:N/A (2023). PubMed: 36625203
- Wang Q et al. A novel drug-like water-soluble small molecule Focal Adhesion Kinase (FAK) activator promotes intestinal mucosal healing. Curr Res Pharmacol Drug Discov 4:100147 (2023). PubMed: 36632414
- Zhong Y & Lan J Overexpression of Eukaryotic translation initiation factor 3D induces stem cell-like properties and metastasis in cervix cancer by activating FAK through inhibiting degradation of GRP78. Bioengineered 13:1952-1961 (2022). WB, IHC-P ; Human . PubMed: 35104170
- Wang J et al. LINC00941 promotes pancreatic cancer malignancy by interacting with ANXA2 and suppressing NEDD4L-mediated degradation of ANXA2. Cell Death Dis 13:718 (2022). PubMed: 35977942