G-LISA Small GTPase Activation Assays offer a fast and sensitive method for performing small G-protein activation assays. Advantages of G-LISA activation assays include:
Information Resources: About, Technical Tips, G-LISA Protocol Video, G-LISA Technical Guide
Related Products: Activators, Proteins, Antibodies, Pull-down Assays
Many publications cite the use of Cytoskeleton's G-LISA kits in the Materials and Methods section. Usually the citation is associated with a particular result in the form of a graph or image that helps the authors present their findings. This indicates the utility of the Kits to produce publication quality data in a short timeframe thus helping improve the productivity of your efforts. Example citations for G-LISA assay kits are shown below. More citations are available on individual product pages.
Nobe et al., 2012. Two distinct dysfunctions in diabetic mouse mesenteric artery contraction are caused by changes in the Rho A–Rho kinase signaling pathway. E. J. Pharmacol. v 683, pp 217-225.
Wang et al., 2012. RhoA/ROCK-dependent moesin phosphorylation regulates AGE-induced endothelial cellular response. Cardiovascular Diabetology. v 11, p 7.
Zuo et al., 2012. Cdc42 negatively regulates intrinsic migration of highly aggressive breast cancer cells. J. Cell. Physiol. v 227, pp 1399-1407.
Alvarez et al. (2010). Failure of Bay K 8644 to induce RhoA kinase-dependent calcium sensitization in rabbit blood vessels. British J of Pharmacology 160 ,1326-37.
Heckman-Stoddard et al. (2009). Haploinsufficiency for p190B RhoGAP inhibits MMTV-Neu tumor progression. Breast Cancer Research 11 ,http://breast-cancer-research.com/content/11/4/R61.
Hammar et al. (2009). Role of the Rho-ROCK (Rho-Associated Kinase) Signaling Pathway in the Regulation of Pancreatic β-Cell Function. Endocrinology 150 ,2072-2079.
Chastre et al. (2009). TRIP6, a novel molecular partner of the MAGI-1 scaffolding molecule, promotes invasiveness. FASEB Journal 23 ,916–928.
Kinoshita et al., 2008. Mol Biol Cell. 19, 2289
Moniz et al. (2008). WNK2 modulates MEK1 activity through the Rho GTPase pathway. Cellular Signalling 20 ,1762-68.
Lesato et al. (2008). Tiotropium Bromide Attenuates Respiratory Syncytial Virus Replication in Epithelial Cells. Respiration 76 ,434-441.
Kinoshita et al. (2008). Apical Accumulation of Rho in the Neural Plate Is Important for Neural Plate Cell Shape Change and Neural Tube Formation. Molecular Biology of the Cell 19 ,2289-2299.
Scott et al., 2007. J Invest Dermatol. v 127, p 668.
Schreibelt et al. (2007). Reactive oxygen species alter brain endothelial tight junction dynamics via RhoA, PI3 kinase, and PKB signaling. FASEB Journal 21 ,3666-3676.
Tanaka et al. (2007). Neural Expression of G Protein-coupled Receptors GPR3, GPR6, and GPR12 Up-regulates Cyclic AMP Levels and Promotes Neurite Outgrowth. J. Biol. Chem 282 ,10506-10515.
Higashibata et al., 2006. BMC Biochem. 7, 19
Zuo et al., 2006. Biochem Biophys Res Commun. 351, 361
Woods and Beier, 2006. J Biol Chem. 281, 13134
Howe and Addison, 2012. RhoB controls endothelial cell morphogenesis in part via negative regulation of RhoA. Vascular Cell. v 4, p 1.
Yang and Kim, 2012. The RhoA-ROCK-PTEN pathway as a molecular switch for anchorage dependent cell behavior. Biomaterials. v 33, pp 2902-2915.
Garrido-Gomez et al., 2012. Annexin A2 is critical for embryo adhesiveness to the human endometrium by RhoA activation through F-actin regulation. FASEB J. doi: 10.1096/fj.12-204008.
Greco et al., 2012. Chemotactic effect of prorenin on human aortic smooth muscle cells: a novel function of the (pro)renin receptor. Cardiovasc Res. doi: 10.1093/cvr/cvs204.
Chen et al., 2012. Inhibition of tumor cell growth, proliferation and migration by X-387, a novel active-site inhibitor of mTOR. Biochem. Pharmacol. v 83, pp 1183-1194.
Zhou et al., 2012. HSV-mediated gene transfer of C3 transferase inhibits Rho to promote axonal regeneration. Exp. Neurol. http://dx.doi.org/10.1016/j.expneurol.2012.06.016.
McCoy et al., 2012. Protease-activated receptor 1 (PAR1) coupling to Gq/11 but not to Gi/o or G12/13 is mediated by discrete amino acids within the receptor second intracellular loop. Cellular Signalling. v 24, pp 1351-1360.
Ramseyer et al., 2012. Tumor Necrosis Factor α Decreases Nitric Oxide Synthase Type 3 Expression Primarily via Rho/Rho Kinase in the Thick Ascending Limb. Hypertension. v 59, pp 1145-1150.
Dhaliwal et al., 2012. Cellular cytoskeleton dynamics modulates non-viral gene delivery through RhoGTPases. PLoS ONE. v 7, e35046.
Jin et al. (2011). Increased SRF Transcriptional Activity is a Novel Signature of Insulin Resistance in Humans and Mice. J Clin Invest.
Ganguly et al. (2011). Adiponectin Increases LPL Activity via RhoA/ROCK-Mediated Actin Remodelling in Adult Rat Cardiomyocytes. Endocrinology 152 ,247.
Rapier et al., 2010. Cancer Cell Int. 10, 24
Nini L, Dagnino L. (2010). Accurate and reproducible measurements of RhoA activation in small samples of primary cells. Anal Biochem 398 ,135-7.
Yang et al. (2010). Fluoride induces vascular contraction through activation of RhoA/Rho kinase pathway in isolated rat aortas. Environmental Toxicology and Pharmacology 29 ,290-296.
Musso et al. (2010). Relevance of the mevalonate biosynthetic pathway in the regulation of bone marrow mesenchymal stromal cell-mediated effects on T cell proliferation and B cell survival. Haematologica DOI: 10.3324/haematol.2010.031633.
Lichtenstein et al. (2010). Secretase-Independent and RhoGTPase/PAK/ERK-Dependent Regulation of Cytoskeleton Dynamics in Astrocytes by NSAIDs and Derivatives. J Alz Dis 22 ,1135.
Ridgway et al. (2010). Modulation of GEF-H1 Induced Signaling by Heparanase in Brain Metastatic Melanoma Cells. J Cellular Biochemistry 111 ,1299-1309.
Fang et al. (2010). Allogeneic Human Mesenchymal Stem Cells Restore Epithelial Protein Permeability in Cultured Human Alveolar Type II Cells by Secretion of Angiopoietin-1. J Biol Chem 285 ,26211-26222.
Romero et al. (2010). Chronic Ethanol Exposure Alters the Levels, Assembly, and Cellular Organization of the Actin Cytoskeleton and Microtubules in Hippocampal Neurons in Primary Culture. Toxicol. Sci. 118 ,602-612.
Rapier et al. (2010). The extracellular matrix microtopography drives critical changes in cellular motility and Rho A activity in colon cancer cells. Cancer Cell International 10 ,24.
Hammar et al. (2009). Role of the Rho-ROCK (Rho-Associated Kinase) Signaling Pathway in the Regulation of Pancreatic β-Cell Function. Endocrinology 150 ,2072-2079.
Chastre et al. (2009). TRIP6, a novel molecular partner of the MAGI-1 scaffolding molecule, promotes invasiveness. FASEB Journal 23 ,916–928.
Ramirez et al., 2008. J Immunol. 180, 1854
Sequeira et al. (2008). Rho GTPases in PC-3 prostate cancer cell morphology, invasion and tumor cell diapedesis. Clinical and Experimental Metastatis 25 ,569-579.
Moore et al. (2008). Rho inhibition recruits DCC to the neuronal plasma membrane and enhances axon chemoattraction to netrin 1. Development 135 ,2855-2864.
Kinoshita et al. (2008). Apical Accumulation of Rho in the Neural Plate Is Important for Neural Plate Cell Shape Change and Neural Tube Formation. Molecular Biology of the Cell 19 ,2289-2299.
Seifert et al. (2008). Differential activation of Rac1 and RhoA in neuroblastoma cell fractions. Neurosci Lett 450 ,176-180.
Korobova and Svitkina (2008). Arp2/3 Complex Is Important for Filopodia Formation, Growth Cone Motility, and Neuritogenesis in Neuronal Cells. Mol. Biol. Cell. 19 ,1561-1574.
Mercer and Helenius (2008). Vaccinia Virus Uses Macropinocytosis and Apoptotic Mimicry to Enter Host Cells. Science 320 ,531.
Keely et al., 2007. Methods Enzymol. v 426, p 27.
Scott et al., 2007. J Invest Dermatol. v 127, p 668.
Schreibelt et al. (2007). Reactive oxygen species alter brain endothelial tight junction dynamics via RhoA, PI3 kinase, and PKB signaling. FASEB Journal 21 ,3666-3676.
Tanaka et al. (2007). Neural Expression of G Protein-coupled Receptors GPR3, GPR6, and GPR12 Up-regulates Cyclic AMP Levels and Promotes Neurite Outgrowth. J. Biol. Chem 282 ,10506-10515.
Bradley et al., 2006. Mol Biol Cell. 17, 4827
Dhaliwal et al., 2012. Cellular cytoskeleton dynamics modulates non-viral gene delivery through RhoGTPases. PLoS ONE. v 7, e35046.
Halpert et al. (2011). Rac-dependent doubling of HeLa cell area and impairment of cell migration and cell cycle by compounds from Iris germanica. Protoplasma DOI: 10.1007/s00709-010-0254-1.
Vives et al. (2011). The Rac1 exchange factor Dock5 is essential for bone resorption by osteoclasts. Journal of Bone and Mineral Research 26 ,1099.
Tanaka et al., 2010. Biochem Biophys Res Commun. v 399, p 677.
Johanna et al. (2010). Rac1 activity changes are associated with neuronal pathology and spatial memory long-term recovery after global cerebral ischemia. Neurochem International 57 ,762-773.
Lichtenstein et al. (2010). Secretase-Independent and RhoGTPase/PAK/ERK-Dependent Regulation of Cytoskeleton Dynamics in Astrocytes by NSAIDs and Derivatives. J Alz Dis 22 ,1135.
Ridgway et al. (2010). Modulation of GEF-H1 Induced Signaling by Heparanase in Brain Metastatic Melanoma Cells. J Cellular Biochemistry 111 ,1299-1309.
Fang et al. (2010). Allogeneic Human Mesenchymal Stem Cells Restore Epithelial Protein Permeability in Cultured Human Alveolar Type II Cells by Secretion of Angiopoietin-1. J Biol Chem 285, 26211-26222.
Romero et al. (2010). Chronic Ethanol Exposure Alters the Levels, Assembly, and Cellular Organization of the Actin Cytoskeleton and Microtubules in Hippocampal Neurons in Primary Culture. Toxicol. Sci. 118 ,602-612.
Baumer et al., 2009. J Cell Physiol. 220, 716
Chastre et al. (2009). TRIP6, a novel molecular partner of the MAGI-1 scaffolding molecule, promotes invasiveness. FASEB Journal 23 ,916–928.
Kawther Abu-Elneel, Tomoyo Ochiishi, Miguel Medina, Monica Remedi, Laura Gastaldi, Alfredo Caceres, and Kenneth S. Kosik (2008). A delta-Catenin Signaling Pathway Leading to dendritic protrusions. J Biol Chem 283 ,32781-32791.
Mercer and Helenius (2008). Vaccinia Virus Uses Macropinocytosis and Apoptotic Mimicry to Enter Host Cells. Science 320 ,531.
Pontow et al., 2007. Virology. 368, 1
Binker et al., 2011. TGF-β1 increases invasiveness of SW1990 cells through Rac1/ROS/NF-κB/IL-6/MMP-2. Biochem. Biophys. Res. Comm. v 405, pp 140-145.
Sayedyahossein et al., 2012. Essential role of integrin-linked kinase in regulation of phagocytosis in keratinocytes. FASEB J. doi: 10.1096/fj.12-207852.
Kikuchi et al., 2012. Protein kinase C iota as a therapeutic target in alveolar rhabdomyosarcoma. Oncogene. doi:10.1038/onc.2012.46.
Eggers et al., 2012. STE20-related Kinase Adaptor Protein α (STRADα) Regulates Cell Polarity and Invasion through PAK1 Signaling in LKB1-null Cells. J. Biol. Chem. v 287, pp 18758-18768.
Vives et al. (2011). The Rac1 exchange factor Dock5 is essential for bone resorption by osteoclasts. Journal of Bone and Mineral Research 26 ,1099.
McHenry et al. (2010). P190B RhoGAP has pro-tumorigenic functions during MMTV-Neu mammary tumorigenesis and metastasis. Breast Cancer Res.
Johanna et al. (2010). Rac1 activity changes are associated with neuronal pathology and spatial memory long-term recovery after global cerebral ischemia. Neurochem International 57 ,762-773.
Baumer et al., 2009. J Cell Physiol. 220, 716
Heckman-Stoddard et al. (2009). Haploinsufficiency for p190B RhoGAP inhibits MMTV-Neu tumor progression. Breast Cancer Research 11 ,http://breast-cancer-research.com/content/11/4/R61.
Chastre et al. (2009). TRIP6, a novel molecular partner of the MAGI-1 scaffolding molecule, promotes invasiveness. FASEB Journal 23 ,916–928.
Ramirez et al., 2008. J Immunol. 180, 1854
Moniz et al. (2008). WNK2 modulates MEK1 activity through the Rho GTPase pathway. Cellular Signalling 20 ,1762-68.
Pontow et al., 2007. Virology. 368, 1
Chen et al., 2012. Inhibition of tumor cell growth, proliferation and migration by X-387, a novel active-site inhibitor of mTOR. Biochem. Pharmacol. v 83, pp 1183-1194.
Eggers et al., 2012. STE20-related Kinase Adaptor Protein α (STRADα) Regulates Cell Polarity and Invasion through PAK1 Signaling in LKB1-null Cells. J. Biol. Chem. v 287, pp 18758-18768.
Dhaliwal et al., 2012. Cellular cytoskeleton dynamics modulates non-viral gene delivery through RhoGTPases. PLoS ONE. v 7, e35046.
Oliver et al., 2011. Br J Cancer. v 104, p 324.
McHenry et al. (2010). P190B RhoGAP has pro-tumorigenic functions during MMTV-Neu mammary tumorigenesis and metastasis. Breast Cancer Res.
Lichtenstein et al. (2010). Secretase-Independent and RhoGTPase/PAK/ERK-Dependent Regulation of Cytoskeleton Dynamics in Astrocytes by NSAIDs and Derivatives. J Alz Dis 22 ,1135.
Schlegel and Waschke (2010). Impaired cAMP and Rac 1 Signaling Contribute to TNF-α-induced Endothelial Barrier Breakdown in Microvascular Endothelium. Microcirculation 16 ,521.
Stankiewicz et al. (2010). GTPase activating protein function of p85 facilitates uptake and recycling of the β1 integrin. Biochemical and Biophysical Research Communications 391 ,443.
Romero et al. (2010). Chronic Ethanol Exposure Alters the Levels, Assembly, and Cellular Organization of the Actin Cytoskeleton and Microtubules in Hippocampal Neurons in Primary Culture. Toxicol. Sci. 118 ,602-612.
Heckman-Stoddard et al. (2009). Haploinsufficiency for p190B RhoGAP inhibits MMTV-Neu tumor progression. Breast Cancer Research 11 ,http://breast-cancer-research.com/content/11/4/R61.
Antonov et al., 2012. Regulation of endothelial barrier function by TGF-β type I receptor ALK5: Potential role of contractile mechanisms and heat shock protein 90. J. Cell. Physiol. v 227, pp 759-771.
Greco et al., 2012. Chemotactic effect of prorenin on human aortic smooth muscle cells: a novel function of the (pro)renin receptor. Cardiovasc Res. doi: 10.1093/cvr/cvs204.
Oshikawa et al., 2012. Novel role of p66Shc in ROS-dependent VEGF signaling and angiogenesis in endothelial cells. Am. J. Physiol. Heart Circ. Physiol. v 302, pp H724-H732.
Chen et al., 2012. Inhibition of tumor cell growth, proliferation and migration by X-387, a novel active-site inhibitor of mTOR. Biochem. Pharmacol. v 83, pp 1183-1194.
Montalvo-Ortiz et al., 2012. Characterization of EHop-016, a novel small molecule inhibitor of Rac GTPase. J. Biol. Chem. v 287, pp 13228-13238.
Stefanini et al., 2012. Rap1-Rac1 Circuits Potentiate Platelet Activation. Arterioscler Thromb Vasc Biol. v 32, pp 434–441.
Vives et al. (2011). The Rac1 exchange factor Dock5 is essential for bone resorption by osteoclasts. Journal of Bone and Mineral Research 26 ,1099.
Gil-Henn et al., 2012. Arg/Abl2 promotes invasion and attenuates proliferation of breast cancer in vivo. Oncogene. doi:10.1038/onc.2012.284
Signal Amplification and OptimizationQuestion 1: How can I increase the difference in Rho activation levels between control samples and stimulated samples? Question 2: How can I eliminate high variability between samples when using the absorbance-based G-LISA assays? Question 3: How do I reduce high background readings when using the luminescence-based kits (Cat. # BK121 and BK126)? Activation Conditions and Cell/Tissue CompatibilityQuestion 4: Will the G-LISA assays work for my specific cell type? Question 5: How can I activate Rho, Rac, or Cdc42 GTPases? Question 6: How should I create a cell extract for testing the G-LISA kits? Question 7: Can I use the same lysis buffer to prepare lysates for use in the RhoA, Rac, and Cdc42 G-LISA assays? Question 8: Can I use the G-LISA kits to measure GTPase activity in tissue lysates?
3-D/3D cultures and G-LISAsQuestion 9: Can G-LISA assays be used with cells grown in matrigel or other 3-D growth conditions?
Detection of GTPase IsoformsQuestion 10: Can I use the G-LISA kits to detect other isoforms of the Rho, Rac, and Ral GTPases?
Equipment RequirementsQuestion 11: Which plate shakers are suitable for the G-LISA assays?
|
G-LISA RhoA Activation Assay Biochem Kit (Colorimetric format)
G-LISA Cdc42 Activation Assay Biochem Kit (Colorimetric format)
G-LISA Cdc42 Activation Assay Biochem Kit (Colorimetric format)
G-LISA Arf1 Activation Assay Biochem Kit (Colorimetric Based)