Anti-FAK (phospho S732) antibody (ab4792)
- Datasheet
- Specific References (4)
- Protocols
Overview
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Product nameAnti-FAK (phospho S732) antibody
See all FAK primary antibodies -
DescriptionRabbit polyclonal to FAK (phospho S732)
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Host speciesRabbit
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SpecificityThe antibody also reacts with FRNK.
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Tested applicationsSuitable for: IHC-P, ICC/IF, WBmore details
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Species reactivityReacts with: Mouse, Rat, Human
Predicted to work with: Chicken, Xenopus laevis
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Immunogen
Synthetic peptide (Human) derived from the region of human Focal Adhesion Kinase that contains serine 732. The sequence is conserved in mouse, rat and chicken.
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Positive control
- Cdk5 +/- and cdk5 -/- mouse brain lysates.
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General notes
Focal Adhesion Kinase is a 125 kDa non-receptor protein tyrosine kinase that plays a key role in signalling by growth factors, extracellular matrix and stress signals. Indeed, 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. Cyclin-dependent kinase-5 (Cdk5), a serine/threonine kinase and an important regulator in brain development, has been found to be essential for phosphorylation at serine 732. In the context of the Focal Adhesion Kinase-Related Non-Kinase (FRNK), the c-terminal non-kinase domain of Focal Adhesion Kinase that acts as a dominant negative inhibitor of Focal Adhesion Kinase signalling, other serine kinases appear to readily phosphorylate this same site. The functional significance of phosphorylation on serine 732 is yet to be determined.
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: 7.30
Preservative: 0.05% Sodium azide
Constituents: PBS, 2.5% 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 enzyme and (ii) a generic serine phosphorylated peptide to remove antibody that is reactive with phosphoserine, irrespective of the sequence. The final product is generated by affinity chromatography using a Focal Adhesion Kinase-derived peptide that is phosphorylated at serine 732.
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Primary antibody notesFocal Adhesion Kinase is a 125 kDa non-receptor protein tyrosine kinase that plays a key role in signalling by growth factors, extracellular matrix and stress signals. Indeed, 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. Cyclin-dependent kinase-5 (Cdk5), a serine/threonine kinase and an important regulator in brain development, has been found to be essential for phosphorylation at serine 732. In the context of the Focal Adhesion Kinase-Related Non-Kinase (FRNK), the c-terminal non-kinase domain of Focal Adhesion Kinase that acts as a dominant negative inhibitor of Focal Adhesion Kinase signalling, other serine kinases appear to readily phosphorylate this same site. The functional significance of phosphorylation on serine 732 is yet to be determined.
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
Applications
Our Abpromise guarantee covers the use of ab4792 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-P | 1/200. | |
| ICC/IF | 1/200. | |
| WB | Use a concentration of 0.1 - 0.5 µg/ml. Predicted molecular weight: 125 kDa. |
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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Immunocytochemistry/ Immunofluorescence - Anti-FAK (phospho S732) antibody (ab4792)This image is courtesy of an abreview submitted by Carl Hobbs, King's College London, United KingdomImmunofluorescence labelling of FAK (phospho S732) with antibody ab4792 on mouse Cor 1 cells. Fixative: Formaldehyde. Permeabilization: Tween 20. Blocking step: 1% BSA for 10 mins @ rt°C. ab4792 used at 1/200 for 2 hours in TBS/BSA/azide/Tween. Secondary antibody: anti Rabbit IgG Alexa Fluor® 594 (1/1000). Nuclear positivity of mitotic cells is observed as well as a weaker punctate cytoplasmic positivity. The submitted composite image shows an intensely stained nucleus (upper image) that is clearly a cell in anaphase (lower image of Hoechst staining). To the right side of the upper image, three nuclei exhibit varying degrees of positivity: perhaps these cells are about to enter/leave the mitotic cycle.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FAK (phospho S732) antibody (ab4792)This image is courtesy of an abreview submitted by Carl Hobbs, King's College London, United KingdomImmunohistochemistical detection of FAK (phospho S732) using antibody (ab4792) on Formaldehyde-fixed paraffin-embedded mosue ovary sections. Antigen retrieval step: Heat mediated; Buffer used: Citric acid pH6; Blocking step: 1% BSA for 10 mins @ rt°C. ab4792 (1/200) incubated for 2 hours in TBS/BSA/azide. Secondary antibody: anti Rabbit IgG conjugated to biotin (1/200). The image shows clear nuclear positivity in the middle secondary follicle specific to mitotic cells.
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Western blot - Anti-FAK (phospho S732) antibody (ab4792)Peptide Competition: Extracts prepared from cdk5 +/- mouse brain lysates were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to nitrocellulose. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4oC, then were incubated with FAK pan antibody (1), or 0.34
µ g/mL ab4792 antibody for two hours at room temperature in a 3% BSA TBST buffer, following prior incubation with: no peptide (2), the phosphopeptide corresponding to ab4792 (3), a generic phosphoserine containing peptide (4), or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and signals were detected using the Tropix WesternStar method.Phosphatase Stripping: Extracts prepared from cdk5 +/- mouse brain lysates were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was treated with lambda phosphatase, then incubated with either FAK pan antibody (
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Western blot - Anti-FAK (phospho S732) antibody (ab4792)Influence of cdk5: Extracts prepared from cdk5 +/- (1, 3) and cdk5 -/- (2, 4) mouse brain lysates were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was incubated with either FAK pan antibody (1, 2), or with 0.34µ g/mL ab4792 antibody (3, 4). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and signals were detected using the Tropix WesternStar method. The data show that cdk5 is essential for phosphorylation of FAK at serine 732. Other kinases appear to phosphorylate FRNK on this site. Influence of cdk5: Extracts prepared from cdk5 +/- (1, 3) and cdk5 -/- (2, 4) mouse brain lysates were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was incubated with either FAK pan antibody (1, 2), or with 0.34 µg/mL ab4792 antibody (3, 4). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and signals were de
References
This product has been referenced in:
- Jacobshagen M et al. The serotonin 6 receptor controls neuronal migration during corticogenesis via a ligand-independent Cdk5-dependent mechanism. Development 141:3370-7 (2014). WB ; Mouse . Read more (PubMed: 25078650) »
- Esencay M et al. CXCR7 is induced by hypoxia and mediates glioma cell migration towards SDF-1a. BMC Cancer 13:347 (2013). WB ; Human . Read more (PubMed: 23865743) »
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