This antibody gave a positive result in ELISA against the immunizing peptide (ab40145). It gave a negative result in ELISA against the non-modified equivalent peptide (ab53601). This indicates that it is specific for the modified peptide.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 119 kDa (predicted molecular weight: 119 kDa).
Use a concentration of 5 µg/ml.
Use at an assay dependent concentration.
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.
Expressed in all organs tested, in lymphoid cell lines, but most abundantly in brain.
Belongs to the protein kinase superfamily. Tyr protein kinase family. FAK subfamily. Contains 1 FERM domain. Contains 1 protein kinase 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.
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.
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab39967 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.
ICC/IF image of ab39967 stained human HeLa cells. The cells were methanol fixed (5 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab39967, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
Western blot - Anti-FAK (phospho Y397) antibody (ab39967)
All lanes : Anti-FAK (phospho Y397) antibody (ab39967) at 1 µg/ml
Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate Lane 2 : NIH 3T3 treated with Vanadate and PDGF Whole Cell Lysate Lane 3 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate with Mouse FAK (phospho Y397) peptide (ab40145) at 1 µg/ml Lane 4 : NIH 3T3 treated with Vanadate and PDGF Whole Cell Lysate with Mouse FAK (phospho Y397) peptide (ab40145) at 1 µg/ml Lane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate with Mouse FAK peptide (ab53601) at 1 µg/ml Lane 6 : NIH 3T3 treated with Vanadate and PDGF Whole Cell Lysate with Mouse FAK peptide (ab53601) at 1 µg/ml
Lysates/proteins at 10 µg per lane.
Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size : 119 kDa Observed band size : 119 kDa
McKay TB et al. Acute hypoxia influences collagen and matrix metalloproteinase expression by human keratoconus cells in vitro. PLoS One12:e0176017 (2017).
Read more (PubMed: 28426715) »
Sero JE & Bakal C Multiparametric Analysis of Cell Shape Demonstrates that ß-PIX Directly Couples YAP Activation to Extracellular Matrix Adhesion. Cell Syst4:84-96.e6 (2017).
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