Key Specifications Table
|Species Reactivity||Key Applications||Host||Format||Antibody Type|
|H, M, R||WB||M||Purified||Monoclonal Antibody|
|Presentation||Purified mouse monoclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.|
|Application||Detect RalB using this Anti-RalB Antibody, clone 25 validated for use in WB.|
|Safety Information according to GHS|
|Storage and Shipping Information|
|Storage Conditions||Stable for 1 year at 2-8ºC from date of receipt.|
|Material Size||100 µg|
Anti-RalB Antibody, clone 25 SDS
|Anti-RalB (clone 25) (Purified Mouse Monoclonal IgG) - DAM1483560||DAM1483560|
|Anti-RalB, clone 25 - 2137084||2137084|
|Anti-RalB, clone 25 - 1947718||1947718|
|Anti-RalB, clone 25 - 2018119||2018119|
|Anti-RalB, clone 25 - 2549986||2549986|
|Anti-RalB, clone 25 - 3589743||3589743|
|Anti-RalB, clone 25 - DAM1745372||DAM1745372|
|Anti-RalB, clone 25 - DAM1776478||DAM1776478|
|Anti-RalB, clone 25 - JBC1900513||JBC1900513|
|Anti-RalB, clone 25 - NG1605292||NG1605292|
|Reference overview||Pub Med ID|
|Discovery and characterization of small molecules that target the GTPase Ral.|
Yan, C; Liu, D; Li, L; Wempe, MF; Guin, S; Khanna, M; Meier, J; Hoffman, B; Owens, C; Wysoczynski, CL; Nitz, MD; Knabe, WE; Ahmed, M; Brautigan, DL; Paschal, BM; Schwartz, MA; Jones, DN; Ross, D; Meroueh, SO; Theodorescu, D
Nature 515 443-7 2014
The Ras-like GTPases RalA and RalB are important drivers of tumour growth and metastasis. Chemicals that block Ral function would be valuable as research tools and for cancer therapeutics. Here we used protein structure analysis and virtual screening to identify drug-like molecules that bind to a site on the GDP-bound form of Ral. The compounds RBC6, RBC8 and RBC10 inhibited the binding of Ral to its effector RALBP1, as well as inhibiting Ral-mediated cell spreading of murine embryonic fibroblasts and anchorage-independent growth of human cancer cell lines. The binding of the RBC8 derivative BQU57 to RalB was confirmed by isothermal titration calorimetry, surface plasmon resonance and (1)H-(15)N transverse relaxation-optimized spectroscopy (TROSY) NMR spectroscopy. RBC8 and BQU57 show selectivity for Ral relative to the GTPases Ras and RhoA and inhibit tumour xenograft growth to a similar extent to the depletion of Ral using RNA interference. Our results show the utility of structure-based discovery for the development of therapeutics for Ral-dependent cancers.
|Genetic Deletion of RALA and RALB Small GTPases Reveals Redundant Functions in Development and Tumorigenesis.|
Pascal Peschard,Afshan McCarthy,Val Leblanc-Dominguez,Maggie Yeo,Sabrina Guichard,Gordon Stamp,Christopher J Marshall
Current biology : CB 22 2012
RAL small GTPases, encoded by the Rala and Ralb genes, are members of the RAS superfamily of small GTPases and can act as downstream effectors of RAS . Although highly similar, distinct functions have been identified for RALA and RALB: RALA has been implicated in epithelial cell polarity , insulin secretion , GLUT4 translocation [4, 5], neurite branching, and neuronal polarity [6, 7], and RALB in tumor cell survival , migration/invasion [9-12], TBK1 activation , and autophagy . To investigate RAL GTPases in
|RalA and RalB proteins are ubiquitinated GTPases, and ubiquitinated RalA increases lipid raft exposure at the plasma membrane.|
Neyraud, V; Aushev, VN; Hatzoglou, A; Meunier, B; Cascone, I; Camonis, J
The Journal of biological chemistry 287 29397-405 2012
Ras GTPases signal by orchestrating a balance among several effector pathways, of which those driven by the GTPases RalA and RalB are essential to Ras oncogenic functions. RalA and RalB share the same effectors but support different aspects of oncogenesis. One example is the importance of active RalA in anchorage-independent growth and membrane raft trafficking. This study has shown a new post-translational modification of Ral GTPases: nondegradative ubiquitination. RalA (but not RalB) ubiquitination increases in anchorage-independent conditions in a caveolin-dependent manner and when lipid rafts are endocytosed. Forcing RalA mono-ubiquitination (by expressing a protein fusion consisting of ubiquitin fused N-terminally to RalA) leads to RalA enrichment at the plasma membrane and increases raft exposure. This study suggests the existence of an ubiquitination/de-ubiquitination cycle superimposed on the GDP/GTP cycle of RalA, involved in the regulation of RalA activity as well as in membrane raft trafficking.
|RalB mobilizes the exocyst to drive cell migration.|
Rossé, Carine, et al.
Mol. Cell. Biol., 26: 727-34 (2006) 2006
The Ras family GTPases RalA and RalB have been defined as central components of the regulatory machinery supporting tumor initiation and progression. Although it is known that Ral proteins mediate oncogenic Ras signaling and physically and functionally interact with vesicle trafficking machinery, their mechanistic contribution to oncogenic transformation is unknown. Here, we have directly evaluated the relative contribution of Ral proteins and Ral effector pathways to cell motility and directional migration. Through loss-of-function analysis, we find that RalA is not limiting for cell migration in normal mammalian epithelial cells. In contrast, RalB and the Sec6/8 complex or exocyst, an immediate downstream Ral effector complex, are required for vectorial cell motility. RalB expression is required for promoting both exocyst assembly and localization to the leading edge of moving cells. We propose that RalB regulation of exocyst function is required for the coordinated delivery of secretory vesicles to the sites of dynamic plasma membrane expansion that specify directional movement.
|RalA but not RalB enhances polarized delivery of membrane proteins to the basolateral surface of epithelial cells.|
Shipitsin, Michail and Feig, Larry A
Mol. Cell. Biol., 24: 5746-56 (2004) 2004
RalA and RalB constitute a family of highly similar (85% identity) Ras-related GTPases. Recently, active forms of both RalA and RalB have been shown to bind to the exocyst complex, implicating them in the regulation of cellular secretion. However, we show here that only active RalA enhances the rate of delivery of E-cadherin and other proteins to their site in the basolateral membrane of MDCK cells, consistent with RalA being a regulator of exocyst function. One reason for this difference is that RalA binds more effectively to the exocyst complex than active RalB does both in vivo and in vitro. Another reason is that active RalA localizes to perinuclear recycling endosomes, where regulation of vesicle sorting is thought to take place, while active RalB does not. Strikingly, analysis of chimeras made between RalA and RalB reveals that high-affinity exocyst binding by RalA is due to unique amino acid sequences in RalA that are distal to the common effector-binding domains shared by RalA and RalB. Moreover, these chimeras show that the perinuclear localization of active RalA is due in part to its unique variable domain near the C terminus. This distinct localization appears to be important for RalA effects on secretion because all RalA mutants tested that failed to localize to the perinuclear region also failed to promote basolateral delivery of E-cadherin. Interestingly, one of these inactive mutants maintained binding to the exocyst complex, suggesting that RalA binding to the exocyst is necessary but not sufficient for RalA to promote basolateral delivery of membrane proteins.
|RAL GTPases are linchpin modulators of human tumour-cell proliferation and survival.|
Chien, Yuchen and White, Michael A
EMBO Rep., 4: 800-6 (2003) 2003
The monomeric RAL (RAS-like) GTPases have been indirectly implicated in mitogenic regulation and cell transformation. Here, we show that RALA and RALB collaborate to maintain tumorigenicity through regulation of both proliferation and survival. Remarkably, this task is divided between these highly homologous isoforms. RALB is specifically required for survival of tumour cells but not normal cells. RALA is dispensable for survival, but is required for anchorage-independent proliferation. Reducing the 'oncogenic burden' in human tumour cells relieves the sensitivity to loss of RALB. These observations establish RAL GTPases as crucial components of the cellular machinery that are exploited by factors that drive oncogenic transformation.
|Calmodulin binds RalA and RalB and is required for the thrombin-induced activation of Ral in human platelets|
Clough, Richard R, et al
J Biol Chem, 277:28972-80 (2002) 2002