Active Rap1 Detection Kit #8818
Product Information
Storage
GDP: Store at -80°C
GST-RalGDS-RBD: Store at -80°C
Lysis/Binding/Wash Buffer: Store at 4°C
Glutathione Resin: Store at 4°C
SDS Sample Buffer: Store at 4°C
Spin Cup and Collection Tubes: Store at RT
Rap1 Rabbit mAb: Store at 4°C
Product Description
Specificity / Sensitivity
Active Rap1 Detection Kit detects endogenous levels of GTP-bound (active) Rap1 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.
Background
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).
Rap1 and Rap2 belong to the Ras subfamily of small GTPases and are activated by a wide variety of stimuli through integrins, receptor tyrosine kinases (RTKs), G protein-coupled receptors (GPCR), death domain associated receptors (DD-R), and ion channels (5,6). Like other small GTPases, Rap activity is stimulated by guanine nucleotide exchange factors (GEFs) and inactivated by GTPase activating proteins (GAPs). A wide variety of Rap GEFs have been identified: C3G connects Rap1 with RTKs through adaptor proteins such as Crk, Epacs (or cAMP-GEFs) transmit signals from cAMP, and CD-GEFs (or CalDAG-GEFs) convey signals from either or both Ca2+ and DAG (5). Rap1 primarily regulates multiple integrin-dependent processes such as morphogenesis, cell-cell adhesion, hematopoiesis, leukocyte migration and tumor invasion (5,6). Rap1 may also regulate proliferation, differentiation and survival through downstream effectors including B-Raf, PI3K, RalGEF and phospholipases (PLCs) (5-8). Rap1 and Rap2 are not fuctionally redundant as they perform overlapping but distinct functions (9). Recent research indicates that Rap2 regulates Dsh subcellular localization and is required for Wnt signaling in early development (10).
- Takai, Y. et al. (2001) Physiol Rev 81, 153-208.
- Colicelli, J. (2004) Sci STKE 2004, RE13.
- Wennerberg, K. et al. (2005) J Cell Sci 118, 843-6.
- Vigil, D. et al. (2010) Nat Rev Cancer 10, 842-57.
- Bos, J.L. et al. (2001) Nat Rev Mol Cell Biol 2, 369-77.
- Caron, E. (2003) J Cell Sci 116, 435-40.
- Song, C. et al. (2002) Oncogene 21, 8105-13.
- Rong, R. et al. (2003) J Biol Chem 278, 52497-503.
- Taira, K. et al. (2004) J Biol Chem 279, 49488-96.
- Choi, S.C. and Han, J.K. (2005) EMBO J 24, 985-96.
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