Key features and details
- Rabbit polyclonal to FAK (phospho Y861)
- Suitable for: WB, ICC
- Reacts with: Chicken, Human
- Isotype: IgG
产品名称Anti-FAK (phospho Y861)抗体
参阅全部 FAK 一抗
描述兔多克隆抗体to FAK (phospho Y861)
经测试应用适用于: WB, ICCmore details
种属反应性与反应: Chicken, Human
预测可用于: Rat, Xenopus laevis
Synthetic peptide corresponding to Human FAK (phospho Y861). The sequence is conserved in mouse, rat, chicken and frog.
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.
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存放说明Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Preservative: 0.05% Sodium azide
Constituents: PBS, 50% Glycerol (glycerin, glycerine), 0.1% BSA
BSA is IgG and protease free
Concentration information loading...
纯度Immunogen affinity purified
纯化说明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说明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.
功能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.
组织特异性Expressed in all organs tested, in lymphoid cell lines, but most abundantly in brain.
序列相似性Belongs to the protein kinase superfamily. Tyr protein kinase family. FAK subfamily.
Contains 1 FERM domain.
Contains 1 protein kinase domain.
结构域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.
翻译后修饰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.
细胞定位Cell junction > focal adhesion. Cell membrane. Constituent of focal adhesions.
- Information by UniProt
- FADK 1 antibody
- FADK antibody
- FAK related non kinase polypeptide antibody
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.
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 0.50
µg/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
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.
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.
ab4804 被引用在 3 文献中.
- 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