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You are here:Home » Antibodies » Abs to Protein Kinase Inhibitors » Anti -RhoE (Rho-related GTP Binding Protein, Rnd3)

Anti -RhoE (Rho-related GTP Binding Protein, Rnd3)

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Specifications

Clone Host Grade Applications
Monoclonal Mouse Ascites B IC
The Rho GTPase family member RhoE regulates actin filaments partly by binding to and inhibiting ROCK I, a serine/threonine kinase that induces actomyosin contractility. Here, we show that ROCK I can phosphorylate multiple residues on RhoE in vitro. In cells, ROCK I-phosphorylated RhoE localizes in the cytosol, whereas unphosphorylated RhoE is primarily associated with membranes. Phosphorylation has no effect on RhoE binding to ROCK I, but instead increases RhoE protein stability. Using phospho-specific antibodies, we show that ROCK phosphorylates endogenous RhoE at serine 11 upon cell stimulation with platelet-derived growth factor, and that this phosphorylation requires an active protein kinase C signalling pathway. In addition, we demonstrate that phosphorylation of RhoE correlates with its activity in inducing stress fibre disruption and inhibiting Ras-induced transformation. This is the first demonstration of an endogenous Rho family member being phosphorylated in vivo and indicates that phosphorylation is an important mechanism to control the stability and function of this GTPase-deficient Rho protein.
Applications:
Suitable for use in Western Blot and Immunocytochemistry. Other applications have not been tested.
Catalog #R1998-13
Recommended DilutionWestern Blot: 1:500-1:2000. Detected RhoE/Rnd3 in RIPA lysates from 3T3/A31 cells.
Immunocytochemistry: 1:50. Has been reported by an independent laboratory to show positive immunostaining for RhoE/Rnd3 in 3T3/A31 cells fixed with paraformaldehyde and permeabilized by Triton X-100.
Optimal dilutions to be determined by the researcher.
Included Positive Antigen ControlF4209: Fibroblast Cell, 3T3/A3. Add 2.5ul of 2-mercaptoethanol/100ul of lysate and boil for 5 minutes to reduce the preparation. Load 20ug of reduced lysate per lane for minigels.
Storage and StabilityMay be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Clone TypeMonoclonal
IsotypeIgG
Clone No5H35
HostMouse
SourceMouse
ConcentrationNot Determined
FormSupplied as a liquid, 0.05% sodium azide, before the addition of glycerol to 30%.
PurityAscites
ImmunogenFull length fusion protein corresponding to residues 1-244 of mouse RhoE/Rnd3.
SpecificityRecognizes mouse RhoE/Rnd3, Mr ~29kD. Weakly crossreacts with Rnd1, Mr ~27kD. Species Crossreactivity: human.
Important NoteThis product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.


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