FAK [pY861] Antibody | catalog# XBP-4116
Background
Focal Adhesion Kinase (FAK) is a 125 kDa non-receptor protein tyrosine kinase that is a substrate for Src and a key element in growth factor and integrin signaling. FAK plays a central role in cell spreading, differentiation, migration, cell death and acceleration of the G1 to S phase transition of the cell cycle. Tyrosine 861 of FAK is a major Src phosphorylation site that promotes the binding of FAK to the cytoplasmic tail integrins, and is also involved in cancer.
Description
![FAK [pY861] Antibody](./image/product/00/15/XBP-4116.gif)
Extracts prepared from CEF cells expressing FAK and plated on fibronectin were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4˚C, then were incubated with the FAK [pY861] antibody for two hours at room temperature in a 3% BSA-TBST buffer, 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 HRP conjugate and bands were detected using the Pierce SuperSignal™ method.
Specifications
Formulation
Rabbit polyclonal immunoglobulin in Dulbecco's phosphate buffered saline (without Mg2+ and Ca2+), pH 7.3 (+/- 0.1), 50% glycerol, with 1.0 mg/mL BSA (IgG, protease free) as a carrier.Clonality/Clone
This is a polyclonal antibody.Specificity
Human, mouse and chicken (100% homologous) FAK. Rat and frog (100% homologous) FAK have not been tested, but are expected to react.Source
FAK antibody was produced against a chemically synthesized phosphopeptide derived from the region of human FAK that contains tyrosine 861. The sequence is conserved in mouse, rat, chicken and frog.Purification
FAK antibody was 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 FAK 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 FAK-derived peptide that is phosphorylated at tyrosine 861.Host
FAK [pY861] antibody was raised in rabbit.Please use anti-rabbit secondary antibodies.
Application
For Western blotting applications, we recommend using the antibody at a 1:1000 dilution.Tested Application
WBBuffer
Rabbit polyclonal immunoglobulin in Dulbecco's phosphate buffered saline (without Mg2+ and Ca2+), pH 7.3 (+/- 0.1), 50% glycerol, with 1.0 mg/mL BSA (IgG, protease free) as a carrier.Storage
Store at -20˚C. We recommend a brief centrifugation before opening to settle vial contents. Then, apportion into working aliquots and store at -20˚C. For shipment or short-term storage (up to one week), 2 - 8˚C is sufficient.Positive Control
- Chicken embryo fibroblasts expressing FAK protein and plated on fibronectin.]
Species Reactivity
H, M, C, R, XReference
- Hauck, C.R., et al. (2002) v-Src SH3-enhanced interaction with focal adhesion kinase at beta 1 integrin-containing invadopodia promotes cell invasion. J. Biol. Chem. 277(15):12487-12490.
- Lacalle, R.A., et al. (2002) Specific SHP-2 partitioning in raft domains triggers integrin-mediated signaling via Rho activation. J. Cell Biol. 157(2):277-289.
- Rigacci, S., et al. (2002) Low Mr phosphotyrosine protein phosphatase associates and dephosphorylates p125 focal adhesion kinase, interfering with cell motility and spreading. J. Biol. Chem. 277(44):41631-41636.
- atta, A., et al. (2001) Transformation of chicken embryo fibroblasts by v-src uncouples ?1 integrin-mediated outside-in but not inside-out signaling. Mol. Cell. Biol. 21(21):7295-7306.]