FAK antibody (pTyr397)

Catalog No. ABIN968643

This anti-FAK antibody is a Mouse Monoclonal antibody detecting FAK in WB and BI. Suitable for Human. This Primary Antibody has been cited in 3+ publications.

Quick Overview for FAK antibody (pTyr397) (ABIN968643)

Target: FAK (PTK2) (PTK2 Protein tyrosine Kinase 2 (PTK2))

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This FAK antibody is un-conjugated

Application: Western Blotting (WB), BioImaging (BI)

Clone: 14-FAK

Product Details anti-FAK Antibody

Binding Specificity: pTyr397

Characteristics: 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us for technical protocols.
3. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
6. Triton is a trademark of the Dow Chemical Company.

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Human FAK (pY397) Peptide

Isotype: IgG1

Application Details

Application Notes: Bioimaging
1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and permeabilize the cells using either 90% methanol, or Triton™ X-100: a. Add 100 myl of -20°C 90% methanol to each well and incubate for 5 minutes at RT. OR b. Add 100 myl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.
4. Remove the permeabilization buffer, and wash the wells twice with 100 myl of 1× PBS.
5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 minutes at RT.
6. Remove the blocking buffer and add 50 myl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate for 1 hour at RT.
7. Remove the primary antibody, and wash the wells three times with 100 myl of 1× PBS.
8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl to each well, and incubate in the dark for 1 hour at RT.
9. Remove the second step reagent, and wash the wells three times with 100 myl of 1× PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.

Comment:

Related Products: ABIN968536, ABIN968630, ABIN967389, ABIN967736

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 250 μg/mL

Buffer: Aqueous buffered solution containing BSA, glycerol, and ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: -20 °C

Storage Comment: Store undiluted at -20°C.

References for anti-FAK antibody (ABIN968643)

McLean, Fincham, Frame: "v-Src induces tyrosine phosphorylation of focal adhesion kinase independently of tyrosine 397 and formation of a complex with Src." in: The Journal of biological chemistry, Vol. 275, Issue 30, pp. 23333-9, (2000) (PubMed).

Ruest, Roy, Shi, Mernaugh, Hanks: "Phosphospecific antibodies reveal focal adhesion kinase activation loop phosphorylation in nascent and mature focal adhesions and requirement for the autophosphorylation site." in: Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research, Vol. 11, Issue 1, pp. 41-8, (2000) (PubMed).

Calalb, Zhang, Polte, Hanks: "Focal adhesion kinase tyrosine-861 is a major site of phosphorylation by Src." in: Biochemical and biophysical research communications, Vol. 228, Issue 3, pp. 662-8, (1997) (PubMed).

Target Details for FAK

Target: FAK (PTK2) (PTK2 Protein tyrosine Kinase 2 (PTK2))

Alternative Name: FAK

Background: Focal Adhesion Kinase (FAK) is a cytoplasmic tyrosine kinase that colocalizes with integrins in focal adhesions. This cellular localization is directed by a 125 amino acid sequence at the C-terminus called the Focal Adhesion Targeting sequence (FAT). The binding of extracellular matrix ligands to integrins triggers autophosphorylation at Tyr-397, and activation of FAK through phosphorylation of Tyr residues (Tyr-576 and Tyr577) in the kinase domain activation loop. For example, cell adhesion to a fibronectin substratum involves concurrent activation of Src and phosphorylation of the FAK activation loop. In addition, phosphorylation of other Tyr residues (Tyr-925, and Tyr-861) creates binding sites for SH2 domains of intracellular signaling molecules such as Src, PI3 kinase, and Grb2. FAK's ability to bind numerous structural and signaling proteins via a variety of interactions is important for FAK activation level, and for FAK interaction with a variety of substrates localized to sites of cell adhesion. Thus, FAK activity is regulated by a complex set of phosphorylation sites, and this phospho-regulation could be important for cell motility, cell growth, cytoskeletal organization, and adhesion-dependent cell survival.
Synonyms: Focal Adhesion Kinase (pY397)

Molecular Weight: 116-125 kDa

Pathways: Response to Growth Hormone Stimulus, CXCR4-mediated Signaling Events, Smooth Muscle Cell Migration, Signaling of Hepatocyte Growth Factor Receptor, VEGF Signaling