Catalog # FP3481

FHOD1 (Thr-1141), phospho-specific Antibody

Rabbit Polyclonal

Application / Dilution

ELISA	1:2000
WB	1:1000

Size 100 μl

Species Reactivity Hu, Rt, Ms

MW 140 kDa

Formins include several families of proteins that regulate actin cytoskeletal dynamics via two conserved formin homology domains, FH1 and FH2. The FH1 region contains poly-proline stretches that promote interactions with profilin. The FH2 domain, located C-terminally to the FH1 domain, is highly conserved in formin proteins and possesses actin nucleation and polymerization activities. Through cooperation of FH1 and FH2, formins construct actin-based structures comprising linear, unbranched filaments that are used in stress fibers, actin cables, microspikes, and contractile rings. Several mammalian formins, including mDia1, FRL, and formin homology domain protein 1 (FHOD1) are inhibited through an intramolecular interaction between the C-terminal Dia autoregulatory domain (DAD) and its recognition region at the N-terminus. In FHOD1, this autoinhibitory interaction is disrupted through phosphorylation of Ser-1131, Ser-1137, and Thr-1141 by ROCK. Subsequent FHOD1 activation leads to stress fiber formation. In endothelial cells, thrombin activates this ROCK pathway, leading to FHOD1-mediated stress fiber formation.

References

Takeya, R. et al. (2008) EMBOJ 27:618.
Takeya, R. & Sumimoto, H. (2003) J Cell Sci. 116:4567.
Westendorf, J.J. (2001) J Biol Chem. 276:46453.

Western blot of FHOD1 phosphorylation in human K562 cells stimulated with calyculin A (100 nM) for 30 min. (lanes 1 & 3). The blot was then treated with lambda phosphatase (lanes 2 & 4). Blots were probed with mouse polyclonal anti-FHOD1 (lanes 1 & 2) and anti-FHOD1 (Thr-1141), phospho-specific antibody (lanes 3 & 4) .

Phospho-FHOD1 (Thr-1141) synthetic peptide (coupled to KLH) corresponding to amino acid residues surrounding Thr-1141 in human FHOD1. This peptide sequence is highly conserved in rat and mouse FHOD1, and is well conserved in FHOD3 (Thr-1399).

*For more information, see UniProt Accession Q9Y613

Rabbit polyclonal, affinity-purified antibody is supplied in 100μl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store 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.

This antibody was cross-adsorbed to unphosphorylated FHOD1 (Thr-1141) peptide before affinity purification using phospho-FHOD1 (Thr-1141) peptide (without carrier). The antibody detects a 140 kDa* protein corresponding to the apparent molecular mass of FHOD1 on SDS-PAGE immunoblots of human K562 and mouse C2C12 cells treated with calyculin A.

*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:

Boddy, KC et al. (2018) Cell Microbiol. Oct; 20(10): e12866. (WB: Hela cells)
Pandey, P. et al. (2018) Dev Cell. 45(5):661. (WB: rat cardiomyocytes)
Dwyer, J. wt al. (2014)Anat Rec (Hoboken). 297(9):1560 (WB: mouse heart, soleus, tibialis anterior)
Iskratsch, T. et al. (2013) Dev Cell. 27(5):545. (WB: mouse MEF, SYF, transfectants)
Truong, D. et al. (2013) Cell Microbiol. 15(12):2051. (WB, ICC: HeLa transfection)
Staus, D.P. et al. (2011) Arterioscler Thromb Vasc Biol. 31(2):360. (WB: mouse, smooth muscle, siRNA)

This kit contains: