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To Purchase # 8820S

8820S 1 Kit (30 rxns) $719.00.0


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Product Includes Quantity Storage Temp
GST-Rhotekin-RBD 4 mg -80°C
GDP 50 µl -80°C
GTP γS 50 µl -80°C
Rho Rabbit Antibody 75 µl -20°C
Glutathione Resin 3 ml 4°C
SDS Sample Buffer 1.5 ml 4°C
Lysis/Binding/Wash Buffer 100 ml 4°C
Spin Cup and Collection Tubes 30 ea 4°C

Product Description

The Active Rho Detection Kit provides all reagents necessary for measuring activation of Rho GTPase in the cell. GST-Rhotekin-RBD fusion protein is used to bind the activated form of GTP-bound Rho, which can then be immunoprecipitated with glutathione resin. Rho activation levels are then determined by western blot using a Rho Rabbit Antibody.

Specificity / Sensitivity

Active Rho Detection Kit detects endogenous levels of GTP-bound (active) Rho as shown in Figure 1. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Species Reactivity: Human, Mouse

The Ras superfamily of small GTP-binding proteins (G proteins) comprise a large class of proteins (over 150 members) that can be classified into at least five families based on their sequence and functional similarities: Ras, Rho, Rab, Arf, and Ran (1-3). These small G proteins have both GDP/GTP-binding and GTPase activities and function as binary switches in diverse cellular and developmental events that include cell cycle progression, cell survival, actin cytoskeletal organization, cell polarity and movement, and vesicular and nuclear transport (1). An upstream signal stimulates the dissociation of GDP from the GDP-bound form (inactive), which leads to the binding of GTP and formation of the GTP-bound form (active). The activated G protein then goes through a conformational change in its downstream effector-binding region, leading to the binding and regulation of downstream effectors. This activation can be switched off by the intrinsic GTPase activity, which hydrolyzes GTP to GDP and releases the downstream effectors. These intrinsic guanine nucleotide exchange and GTP hydrolysis activities of Ras superfamily proteins are also regulated by guanine nucleotide exchange factors (GEFs) that promote formation of the active GTP-bound form and GTPase activating proteins (GAPs) that return the GTPase to its GDP-bound inactive form (4).

Rho family small GTPases, including Rho, Rac, and cdc42, act as molecular switches, regulating processes such as cell migration, adhesion, proliferation, and differentiation. They are activated by guanine nucleotide exchange factors (GEFs), which catalyze the exchange of bound GDP for GTP, and inhibited by GTPase activating proteins (GAPs), which catalyze the hydrolysis of GTP to GDP. A third level of regulation is provided by the stoichiometric binding of Rho GDP dissociation inhibitor (RhoGDI) (5). RhoA, RhoB and RhoC are highly homologous, but appear to have divergent biological functions. Carboxy-terminal modifications and differences in subcellular localization allow these three proteins to respond to and act on distinct signaling molecules (6,7).

1.  Takai, Y. et al. (2001) Physiol Rev 81, 153-208.

2.  Colicelli, J. (2004) Sci STKE 2004, RE13.

3.  Wennerberg, K. et al. (2005) J Cell Sci 118, 843-6.

4.  Vigil, D. et al. (2010) Nat Rev Cancer 10, 842-57.

5.  DerMardirossian, C. and Bokoch, G.M. (2005) Trends Cell Biol. 15, 356-363.

6.  Wennerberg, K. and Der, C.J. (2004) J. Cell Sci. 117, 1301-1312.

7.  Wheeler, A.P. and Ridley, A.J. (2004) Exp Cell Res 301, 43-9.

Entrez-Gene Id 387
Swiss-Prot Acc. P61586

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Active Rho Detection Kit