Catalog # TK6470

Talin Phospho-Regulation Antibody Kit

Antibody Sampler Kit



Talin is an important cytoskeletal component of integrin adhesion sites. Calpins cleave talin precursor (240 kDa) into an amino-terminal globular head domain of 47 kDa and a carboxyl-terminal 190 kDa rod domain. The talin head domain contains a FERM domain, that binds integrins, PIP kinase (Type I), and FAK. The rod domain has several vinculin-binding sites, a second integrin-binding site, and two actin-binding sites. These talin protein-protein interactions are critical for integrin activation, focal adhesion formation and cell migration. Talin regulation may occur through phosphorylation and regulated degradation. The talin head domain binds Smurf1, an E3 ubiquitin ligase, and this interaction leads to talin head ubiquitylation and degradation. Cdk5 can phosphorylate Ser-425 in the head domain, and this inhibits both binding to Smurf1 and subsequent degradation. The S425A talin mutant resists Cdk5 phosphorylation, increases susceptibility to Smurf1-mediated ubiquitylation, and inhibits cell migration. Thus, talin head phosphorylation may be important for regulating adhesion stability and cell migration.

Ratnikov, B. et al. (2005) J. Cell Sci. 118:4921.
Huang, C. et al. (2009) Nat Cell Biol. 11(5):624.
Moser, M. et al. (2009) Science. 324(5929):895.

Western blot of rat PC12 cells stimulated with Calyculin A (100 nM) for 30 min (lanes 1-4). The blot was treated with lambda phosphatase (lanes 2 & 4), then probed with mouse monoclonal anti-Talin (8D4) (lanes 1 & 2) or rabbit polyclonal anti-Talin (Ser-425) (lanes 3 & 4) antibodies.

Immunocytochemical labeling of Talin relative to F-actin in chick fibroblasts. The cells were labeled with mouse monoclonal Talin (Rod domain) antibody (TM4081), then the antibody was detected using appropriate secondary antibody (Red). This labeling is compared to F-actin staining (Green). (Image provided by Dr. Gianluca Gallo at Drexel University).

Rabbit polyclonal and mouse monoclonal antibodies are supplied in phosphate-buffered saline (PBS), 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. The secondary reagents are supplied in the same buffer without azide, and the peptide is supplied in PBS and 0.05% sodium azide. Store all at –20°C. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

Product References:

TP4171 Jin, J.K. et al. (2015) Oncogene. 34(14): 1811. WB: metastatic PCa cells, PC3-MM2, shRNA
TX4175 Jin, J.K. et al. (2015) Oncogene. 34(14): 1811. WB: metastatic PCa cells, PC3-MM2, shRNA
RS3251 Kawasaki, H. et al. (2013) World J Gastroenter. 19(17):2629. WB, ICC: mouse intestinal myofibroblasts
MS3001 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot
RS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot

This kit contains:

Talin (Rod domain) Mouse mAb50 μlWB, E, IP, ICCHu, Rt, Ms, Ck, F240
Talin (Ser-425), phospho-specific Rabbit pAb50 μlWB, E, ICCHu, Rt, Ms, Ck, F240
phospho-Talin (Ser-425) Peptide 50 μgAB, E
Anti-Mouse Ig:HRP Donkey pAb100 μlWB, EMs
Anti-Rabbit Ig Light-Chain Specific:HRP Mouse mAb100 μlWB, E, ICC, IHCRb

The talin phospho-regulation antibody sampler kit can be used to detect the level of phosphorylation of talin (Ser-425) relative to the total expression level of talin. The kit also includes phosho-talin (Ser-425) peptide for antibody blocking experiments and secondary reagents for antibody detection.