Phospho-FAK (Y397) Recombinant Rabbit Monoclonal Antibody [SC54-07]
cat.: ET1610-34
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, IF-Tissue, IHC-P |
| Clonality: | Monoclonal |
| Clone number: | SC54-07 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term. |
| Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 119 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Tyr397 of human FAK. |
| Positive control: | C6 (Rat glioma cell) cell lysates, NIH/3T3, C6, Hela, mouse spleen tissue, rat spleen tissue. |
| Subcellular location: | Cytoplasm, Nucleus, Cell membrane, Cell junction. |
| Recommended Dilutions:
WB IF-Cell IF-Tissue IHC-P |
1:2,000 1:50-1:100 1:50-1:100 1:200 |
| Uniprot #: | SwissProt: Q05397 Human | P34152 Mouse | O35346 Rat |
| Alternative names: | FADK 1 FADK FAK related non kinase polypeptide FAK1 FAK1_HUMAN Focal adhesion kinase 1 Focal adhesion Kinase Focal adhesion kinase isoform FAK Del33 Focal adhesion kinase related nonkinase FRNK p125FAK pp125FAK PPP1R71 Protein phosphatase 1 regulatory subunit 71 Protein tyrosine kinase 2 Protein-tyrosine kinase 2 Ptk2 PTK2 protein tyrosine kinase 2 |
Images
|
Fig1:
Western blot analysis of Phospho-FAK (Y397) on C6 (Rat glioma cell) cell lysate with Rabbit anti-Phospho-FAK (Y397) antibody (ET1610-34) 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: ET1610-34, 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 |
|
Fig2:
Immunocytochemistry analysis of NIH/3T3 cells labeling Phospho-FAK (Y397) with Rabbit anti-Phospho-FAK (Y397) antibody (ET1610-34) 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 (ET1610-34) 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. |
|
Fig3:
Immunocytochemistry analysis of C6 cells labeling Phospho-FAK (Y397) with Rabbit anti-Phospho-FAK (Y397) antibody (ET1610-34) 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 (ET1610-34) 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. |
|
Fig4: 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 (ET1610-34, 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). |
|
Fig5:
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-Phospho-FAK (Y397) antibody (ET1610-34) 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 (ET1610-34) 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. |
|
Fig6:
Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-Phospho-FAK (Y397) antibody (ET1610-34) 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 (ET1610-34) 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. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.