Catalog # RL7421

RhoA + PKA Phosphorylated


Application / Dilution
WB10 μl/lane

Size 100 μl



Rho (A, B, & C) proteins are members of the Ras superfamily of GTPases and 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.

Human recombinant RhoA full length protein (28 kDa) with an N-terminal His-tag was phosphorylated by Protein Kinase A (PKA) in an in vitro kinase assay. The phosphorylated RhoA protein is detected by rabbit polyclonal anti-RhoA (Ser-188) phospho-specific antibody (Cat.#RP1361).

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).

RhoA recombinant protein lysate is supplied at a concentration of 5 ng/µl in electrophoresis sample buffer (62.5 mM Tris pH 6.8, 2% SDS, 5% glycerol, 0.003% bromophenol blue, 0.9% β-mercaptoethanol). Store at –20°C. Do not boil or dilute. 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.

This kit contains: