Anti-FAK (phospho Y861) antibody (ab4804)
Key features and details
- Rabbit polyclonal to FAK (phospho Y861)
- Suitable for: WB
- Reacts with: Chicken
- Isotype: IgG
Overview
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Product name
Anti-FAK (phospho Y861) antibody
See all FAK primary antibodies -
Description
Rabbit polyclonal to FAK (phospho Y861) -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species WB Chicken
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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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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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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
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 0.50
µ 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.
