Catalog # RX1365

phospho-RhoA (Ser-188) Peptide

Blocking Peptide

Application / Dilution

Blocking	1:1000
ELISA	50 ng/well

Size 50 μg

Rho (A, B, & C) proteins are members of the Ras superfamily of GTPases. These proteins regulate a variety of cellular functions, including cell cycle progression, cytoskeletal rearrangement, and gene expression. Rho cycles between the active GTP-bound form and an inactive GDP-bound form. Interconversion between these forms is controlled by guanine nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs). The Rho proteins RhoA, RhoB, and RhoC are highly homologous and contain the consensus amino acid sequences necessary for GDP/GTP-binding and GTPase activity. Post-translational regulation of Rho activity has been shown specifically for RhoA. This Rho protein is phosphorylated in vitro on serine 188 by cAMP- and cGMP-dependent kinases (PKA and PKG). Ser-188 phosphorylation enhances RhoGDI binding and inhibits RhoA-mediated stress fiber formation. Thus, Ser-188 is an important site for negative regulation of RhoA activity.

References

Ellerbroek, S.M. et al. (2003) J. Biol. Chem. 278(21):19023.
Sawada, N. et al. (2001) Biochem. Biophys. Res. Commun. 280:798.
Dong, J.M. et al. (1998) J. Biol. Chem. 273(35):22554.
Lang, P. et al. (1996) EMBO J. 15:510.

Western blot analysis of human RhoA GST fusion recombinant unphosphorylated (lanes 1 & 3) or phosphorylated with PKA (lanes 2, 4, 5 & 6). The blots were probed with anti-Rho (RP1501; lanes 1 & 2) or with anti-RhoA (Ser-188) (RP1361; lanes 3-6). The latter antibody was used in the presence of no peptide (lanes 3 & 4), phospho-RhoA (Ser-188) peptide (lane 5), or a non-specific phosphoserine peptide (lane 6).

Phospho-RhoA (Ser-188) synthetic peptide corresponds to amino acid residues around serine 188 of human RhoA. This peptide sequence is highly conserved in rat and mouse RhoA, but is not found in RhoB or RhoC proteins.

*For more information, see UniProt Accession P61586

Blocking Peptide is supplied in 50µl phosphate-buffered saline 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.

The peptide is specifically recognized by anti-RhoA (Ser-188) phospho-specific antibody (RP1361) in ELISA, and has been shown to block the reactivity of RP1361 during Western blot. In addition, the peptide is recommended for use in blocking RP1361 reactivity in immunocytochemistry.

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

This kit contains: