Analysis Note

ControlIncluded Positive Antigen Control: Catalog #12-305, 3T3/A31 cell lysate. Add 2.5 µL of 2-mercaptoethanol/100 µL of lysate and boil for 5 minutes to reduce the preparation. Load 20 µg of reduced lysate per lane for minigels.

Application

Research CategorySignaling

Immunocytochemistry:
A 1:50 dilution of this antibody 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 (Riento, K., 2003).

Anti-RhoE/Rnd3 Antibody, clone 4 is an antibody against RhoE/Rnd3 for use in IC & WB.

Research Sub CategoryCytoskeletal Signaling

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

RhoE, also known as Rnd3, (Rho family GTPase 3) is a small G-protein that is a member of the Rnd family of Rho GTPases, which is a part of the Ras superfamily. The activity of Rnd1, Rnd2, and Rnd3/RhoE are not activated by the traditional conformational change of the GTP binding like other Rho family members. As the name dictates, Rnd proteins were associated with making a cell “round”. Like other small G-proteins, the do bind GTP, but unlike other G-proteins, they do not hydrolyze GTP to GDP. The actions of RhoE are associated with the actin cytoskeletal organization in response to external stimuli. RhoE acts antagonistically to RhoA in inducing loss of stress fiber formation, in part by binding to the RhoA downstream kinase Rho-associated kinase (ROCK1). This prevents ROCK1 activation and inhibits downstream signaling. RhoE does go through the post-translation modification of farnylation which enables it to localize and bind to various internal membranes. RhoE is also known to effect cycle progression.

Immunogen

Full length fusion protein corresponding to residues 1-244 of mouse RhoE/Rnd3. Clone 4.

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Mouse monoclonal ascites IgG in buffer containing 0.05% sodium azide before the addition of glycerol to 30%. Liquid at -20°C.

Unpurified

Quality

Routinely evaluated by western blot on RIPA lysates from 3T3/A31 cells.

Western Blot Analysis:
A 1:500-1:2000 dilution of this lot detected RhoE/Rnd3 in RIPA lysates from 3T3/A31 cells.

Specificity

This antibody recognizes RhoE/Rnd3, Mr 29 kDa. Weakly cross-reacts with Rnd1, Mr 27 kDa.

Storage and Stability

Stable for 1 year at -20°C from date of receipt. For maximum recovery of product, centrifuge the vial prior to removing the cap.

Target description

RhoE/Rnd3 (~29 kDa); Rnd1 (~27 kDa).