- Datasheet
- Specific References (3)
- Protocols
Overview
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Product nameAnti-FAK (phospho Y861) antibody
See all FAK primary antibodies -
DescriptionRabbit polyclonal to FAK (phospho Y861)
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Host speciesRabbit
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Tested applicationsSuitable for: IHC-Fr, WB, ICC/IFmore details
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Species reactivityReacts with: Mouse, Chicken, Human
Predicted to work with: Rat, Xenopus laevis
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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.
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Positive control
- Chicken embryo fibroblasts expressing Focal Adhesion Kinase protein and plated on fibronectin.
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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
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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Storage bufferpH: 7.40
Preservative: 0.05% Sodium azide
Constituents: PBS, 50% Glycerol, 0.1% BSA
BSA is IgG and protease free -
Concentration information loading... -
PurityImmunogen affinity purified
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Purification notesPurified 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.
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Primary antibody notesFocal 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.
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
Applications
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.
| Application | Abreviews | Notes |
|---|---|---|
| IHC-Fr | Use at an assay dependent concentration. | |
| WB | Use at an assay dependent concentration. Predicted molecular weight: 125 kDa.Can be blocked with FAK (phospho Y861) peptide (ab5304). Can be blocked with FAK peptide - phospho Y861 (phospho and non-phospho pair) or CEF whole cell lysate, positive / negative for Focal Adhesion Kinase (pair). | |
| ICC/IF | Use at an assay dependent concentration. PubMed: 21525268 |
Target
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FunctionNon-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.
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Tissue specificityExpressed in all organs tested, in lymphoid cell lines, but most abundantly in brain.
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Sequence similaritiesBelongs to the protein kinase superfamily. Tyr protein kinase family. FAK subfamily.
Contains 1 FERM domain.
Contains 1 protein kinase domain. -
DomainThe 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 localizationCell junction > focal adhesion. Cell membrane. Constituent of focal adhesions.
- Information by UniProt
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Database links
- Entrez Gene: 396416 Chicken
- Entrez Gene: 5747 Human
- Entrez Gene: 14083 Mouse
- Entrez Gene: 25614 Rat
- Entrez Gene: 399286 Xenopus laevis
- Omim: 600758 Human
- SwissProt: Q00944 Chicken
- SwissProt: Q05397 Human
see all -
Alternative names
- FADK 1 antibody
- FADK antibody
- FAK related non kinase polypeptide antibody
see all
Images
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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 0.50µ g/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)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: 125 kDa
Observed band size: 125 kDa
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.
References
This product has been referenced in:
- 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). Read more (PubMed: 25471072) »
- Ding LW et al. LNK (SH2B3): paradoxical effects in ovarian cancer. Oncogene N/A:N/A (2014). Read more (PubMed: 24704825) »
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