Anti-FAK (phospho Y861) antibody (ab4804)
Key features and details
- Rabbit polyclonal to FAK (phospho Y861)
- Suitable for: WB, ICC
- Reacts with: Chicken
- Isotype: IgG
Overview
-
Product name
Anti-FAK (phospho Y861) antibody
See all FAK primary antibodies -
Description
Rabbit polyclonal to FAK (phospho Y861) -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICCmore details -
Species reactivity
Reacts with: Chicken
Predicted to work with: Rat, Xenopus laevis
-
Immunogen
Synthetic peptide corresponding to Human FAK (phospho Y861). The sequence is conserved in mouse, rat, chicken and frog.
-
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.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.3
Preservative: 0.05% Sodium azide
Constituents: PBS, 50% Glycerol (glycerin, glycerine), 0.1% BSA
BSA is IgG and protease free -
Concentration information loading... -
Purity
Immunogen affinity purified -
Purification notes
Purified 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. -
Primary antibody 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. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
Applications
The Abpromise guarantee
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 |
|---|---|---|
| WB | (2) |
1/1000. Predicted molecular weight: 119 kDa.
|
| ICC |
1/250.
|
| Notes |
|---|
|
WB
1/1000. Predicted molecular weight: 119 kDa. |
|
ICC
1/250. |
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
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
-
Database links
- Entrez Gene: 396416 Chicken
- Entrez Gene: 25614 Rat
- Entrez Gene: 399286 Xenopus laevis
- SwissProt: Q00944 Chicken
- SwissProt: O35346 Rat
- SwissProt: Q91738 Xenopus laevis
- Unigene: 2809 Rat
- Unigene: 6819 Xenopus laevis
-
Alternative names
- FADK 1 antibody
- FADK antibody
- FAK related non kinase polypeptide antibody
see all
Images
-
Immunocytochemistry - Anti-FAK (phospho Y861) antibody (ab4804)
Immunofluorescence analysis of FAK [pY861] was done on 70% confluent log phase A-549 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labelled with ab4804 at 1:250 dilution in 1% BSA and incubated for 3 hours at room temperature and then labelled with a Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with DAPI. F-actin (Panel c: red) was stained with Rhodamine Phalloidin at a 1:300 dilution. Panel d is a merged image showing localization of target protein at focal adhesions. Panel e is a no primary antibody control. The images were captured at 60X magnification.
-
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: 119 kDa
Observed band size: 125 kDa why is the actual band size different from the predicted?
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.
Datasheets and documents
-
Datasheet download
References (3)
ab4804 has been referenced in 3 publications.
- 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). PubMed: 25471072
- Ding LW et al. LNK (SH2B3): paradoxical effects in ovarian cancer. Oncogene N/A:N/A (2014). PubMed: 24704825
- Tan CL et al. Integrin activation promotes axon growth on inhibitory chondroitin sulfate proteoglycans by enhancing integrin signaling. J Neurosci 31:6289-95 (2011). ICC/IF ; Human . PubMed: 21525268
