Phospho-FAK (Y397) Recombinant Rabbit Monoclonal Antibody [SC54-07] - BSA and Azide free
Catalog# HA750212
Phospho-FAK (Y397) Recombinant Rabbit Monoclonal Antibody [SC54-07] - BSA and Azide free
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WB
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IF-Cell
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IF-Tissue
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IHC-P
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Human
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Mouse
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Rat
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ET1610-34
含抗保成分
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unconjugated
Overview
Product Name
Phospho-FAK (Y397) Recombinant Rabbit Monoclonal Antibody [SC54-07] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic phospho-peptide corresponding to residues surrounding Tyr397 of human FAK.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P
Molecular Weight
Predicted band size: 119 kDa
Positive Control
C6 (Rat glioma cell) cell lysates, NIH/3T3, C6, Hela, mouse spleen tissue, rat spleen tissue.
Conjugation
unconjugated
Clone Number
SC54-07
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:2,000
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IF-Cell
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1:50-1:100
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IF-Tissue
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1:50-1:100
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IHC-P
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1:200
Target
Function
Activation of integrins in the extracellular matrix (ECM) of eukaryotic cells promotes the formation of membrane adhesion complexes, known as focal adhesions, which can include cytoskeletal proteins and protein tyrosine kinases, such as focal adhesion kinase (FAK). Phosphorylation events occurring within focal adhesions influence numerous processes that include mitogenic signaling, cell survival, and cell motility. FAK is a non-receptor tyrosine kinase that is ubiquitously expressed and highly conserved between species. FAK is recruited by Integrin clusters and variably phosphorylated depending on the effector molecules present in the focal adhesion. Phosphorylation of FAK Tyr 397 decreases during serum starvation, contact inhibition, and cell cycle arrest, all conditions under which activating FAK Tyr 407 phosphorylation increases.
Background References
1. Kuo SW et al. Regulation of the fate of human mesenchymal stem cells by mechanical and stereo-topographical cues provided by silicon nanowires. Biomaterials 33:5013-22 (2012).
2. Lu H et al. IGFBP2/FAK pathway is causally associated with dasatinib resistance in non-small cell lung cancer cells. Mol Cancer Ther 12:2864-73 (2013).
Sequence Similarity
Belongs to the protein kinase superfamily. Tyr protein kinase family. FAK subfamily.
Tissue Specificity
Detected in B and T-lymphocytes. Isoform 1 and isoform 6 are detected in lung fibroblasts (at protein level). Ubiquitous. Expressed in epithelial cells (at protein level).
Post-translational Modification
Phosphorylated on tyrosine residues upon activation, e.g. upon integrin signaling. Tyr-397 is the major autophosphorylation site, but other kinases can also phosphorylate this residue. Phosphorylation at Tyr-397 promotes interaction with SRC and SRC family members, leading to phosphorylation at Tyr-576, Tyr-577 and at additional tyrosine residues. FGR promotes phosphorylation at Tyr-397 and Tyr-576. FER promotes phosphorylation at Tyr-577, Tyr-861 and Tyr-925, even when cells are not adherent. Tyr-397, Tyr-576 and Ser-722 are phosphorylated only when cells are adherent. Phosphorylation at Tyr-397 is important for interaction with BMX, PIK3R1 and SHC1. Phosphorylation at Tyr-925 is important for interaction with GRB2. Dephosphorylated by PTPN11; PTPN11 is recruited to PTK2 via EPHA2 (tyrosine phosphorylated). Microtubule-induced dephosphorylation at Tyr-397 is crucial for the induction of focal adhesion disassembly; this dephosphorylation could be catalyzed by PTPN11 and regulated by ZFYVE21. Phosphorylation on tyrosine residues is enhanced by NTN1 (By similarity).; Sumoylated; this enhances autophosphorylation.
Subcellular Location
Cytoplasm, Nucleus, Cell membrane, Cell junction.
Synonyms
FADK 1 antibody
FADK antibody
FAK related non kinase polypeptide antibody
FAK1 antibody
FAK1_HUMAN antibody
Focal adhesion kinase 1 antibody
Focal adhesion Kinase antibody
Focal adhesion kinase isoform FAK Del33 antibody
Focal adhesion kinase related nonkinase antibody
FRNK antibody
ExpandImages
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Western blot analysis of Phospho-FAK (Y397) on C6 (Rat glioma cell) cell lysate with Rabbit anti-Phospho-FAK (Y397) antibody (HA750212) at 1/2,000 dilution.
Lysates/proteins at 15 µg/Lane.
Exposure time: 1 minute 4 seconds; ECL: K1802
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA750212, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 119 kDa
Observed band size: 119 kDa -
Immunocytochemistry analysis of NIH/3T3 cells labeling Phospho-FAK (Y397) with Rabbit anti-Phospho-FAK (Y397) antibody (HA750212) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-FAK (Y397) antibody (HA750212) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of C6 cells labeling Phospho-FAK (Y397) with Rabbit anti-Phospho-FAK (Y397) antibody (HA750212) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-FAK (Y397) antibody (HA750212) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
ICC staining of Phospho-FAK (Y397) in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (HA750212, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-Phospho-FAK (Y397) antibody (HA750212) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750212) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-Phospho-FAK (Y397) antibody (HA750212) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750212) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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