Activation Assays

Cytoskeleton's Small G-protein Activation Assays measure the GTP-bound form of the protein from a cell or tissue extract.  

  • Activation assays are available in two formats: the traditional pull-down bead format and our advanced ELISA based G-LISA format.  
  • Measure Arf1Cdc42RacRalRas, and Rho
  • Accurate, sensitive, and user-friendly formats
  • The most trusted and cited activation assays available
  • Watch our video or browse the options below to learn more about which format is right for you. 


G-LISA® is a registered trademark of Cytoskeleton, Inc (CO). All rights reserved.

This table outlines some important considerations when deciding which GTPase activation assay is right for you and summarizes what Cytoskeleton feels will be key decision points when choosing between pull-downs and G-LISAs as a means of measuring activation of small G-proteins.  For more information and help with deciding which of Cytoskeleton’s activation assays are right for you, please check out the informational links on this page or contact our technical support team at tservice@cytoskeleton.com or 303-322-2254, ext 316.

GTPase Activation Assay Selection Guide

Assay Factor

 

Assay Considerations

 

Recommendations

Assay principle

 

Traditional Pull-Down

  • Utilizes affinity beads coated with an effector protein to selectively bind the active GTPase.
  • The signal is analyzed by western blot. 

 

 

Traditional pull-down assays use methods familiar to most labs and therefore provide a familiar point of entry.

 

 

The G-LISA assay is simple to use and has many advantages that should be considered (see below for comparisons).

 

G-LISA

  • Uses a 96 well plate coated with an effector protein to selectively bind the active GTPase.
  • The signal is analyzed by enzyme-linked immunosorbent assay (ELISA).
 

Equipment

 

Traditional Pull-Down

  • SDS-PAGE apparatus
  • Western blot apparatus

 

 

Consider the equipment requirements prior to deciding upon an assay format.

 

 

 

G-LISA

  • Orbital plate shakers (2)
  • Spectrophotometer (set to 490 nm)
  • Multichannel pipette (recommended if handling high number of sample)
 

Starting material required per assay

 

Traditional Pull-Down

300 - 2000 µg per assay

 

 

If starting material is limited, e.g. primary cells, the G-LISA assay is required.

 

G-LISA

5 – 50 µg per assay

 

Cost

 

Traditional Pull-Down

Available as:

  • Combo kit (10 assays each RhoA/Rac1/Cdc42)
  • Starter kit (20 assays)
  • Regular kit (50-80 assays depending on target protein)

 

 

Cost issues to consider:

 

Cost per assay: The G-LISA is the best value per assay.

 

Cost per kit: The traditional pull-down assay starter kits generally provide the lowest cost per kit.  NOTE: Consider the combo kit as a cost effective choice.

 

 

G-LISA

Available in 96 well format. Each well can be broken off and used individually.

 

Sample handling

 

Traditional Pull-Down

Centrifugation steps and western blot analysis makes this assay most suitable for low sample numbers (≤10).

 

 

<10 samples: either assay

>10 samples: G-LISA recommended

NOTE: Dose responses and time courses are often recommended and will greatly increase the number of samples being analyzed.

 

G-LISA

Amenable to high sample numbers (>10) or dose response/time-course analyses. Wells can be detached and used individually.

 

Quantification of results

 

Traditional Pull-Down

Western blot quantification has a very narrow linear range.  Also, multiple manipulations of the samples result in a somewhat variable assay.

 

 

G-LISA assays are recommended for simple, reproducible quantification of results.

 

G-LISA

Provides a simple numeric readout

 

Reagents required that are not in the kits

 

Traditional Pull-Down

  • Cell or tissue lysate
  • Protein quantitation reagent
  • Anti-mouse secondary antibody
  • SDS-PAGE buffers
  • SDS-PAGE gels
  • Molecular weight markers
  • Western blot buffers
  • Western blot detection reagents

 

 

The G-LISA assay provides all reagents necessary to perform 96 activation assays. Wells can be detached and used individually.

 

 

 

G-LISA

  • Cell or tissue lysate
 

 

For more detailed information pertaining to Cytoskeletons Activation Assays, please click on the links below

More about traditional pull-down activation assays

More about G-LISA activation assays

Cytoskeleton's Activation Assays have been cited hundreds of times over the past two decades. Example citations for RhoA G-LISA kit, BK124, is shown, but more citations are available on individual product pages. Please use the "Citations" tab on each individual product page.

AuthorTitleJournalYearArticle Link
Swaminathan, Bhairavi et al.Endothelial Notch signaling directly regulates the small GTPase RND1 to facilitate Notch suppression of endothelial migrationScientific Reports2022ISSN 2045-2322
Nath, Anu S. et al.Modulation of the cell membrane lipid milieu by peroxisomal β-oxidation induces Rho1 signaling to trigger inflammatory responsesCell Reports2022ISSN 2211-1247
Shoda, Tetsuo et al.Desmoplakin and periplakin genetically and functionally contribute to eosinophilic esophagitisNature Communications2021ISSN 2041-1723
Zhou, Qun et al.Inflammatory Immune Cytokine TNF-α Modulates Ezrin Protein Activation via FAK/RhoA Signaling Pathway in PMVECs HyperpermeabilityFrontiers in Pharmacology2021ISSN 1663-9812
Mao, De Yu et al.CLIC1 and CLIC4 mediate endothelial S1P receptor signaling to facilitate Rac1 and RhoA activity and functionScience Signaling2021ISSN 1937-9145
Ma, Yuanyuan et al.Ror2-mediated non-canonical Wnt signaling regulates Cdc42 and cell proliferation during tooth root developmentDevelopment (Cambridge)2021ISSN 1477-9129
Gurusamy, Malarvizhi et al.G-protein-coupled receptor P2Y10 facilitates chemokine-induced CD4 T cell migration through autocrine/paracrine mediatorsNature Communications2021ISSN 2041-1723
Jozic, Ivan et al.Glucocorticoid-mediated induction of caveolin-1 disrupts cytoskeletal organization, inhibits cell migration and re-epithelialization of non-healing woundsCommunications Biology2021ISSN 2399-3642
Rong, Zhouyi et al.Activation of FAK/Rac1/Cdc42-GTPase signaling ameliorates impaired microglial migration response to Aβ42 in triggering receptor expressed on myeloid cells 2 loss-of-function murine modelsFASEB Journal2020ISSN 1530-6860
Lachowski, Dariusz et al.G Protein-Coupled Estrogen Receptor Regulates Actin Cytoskeleton Dynamics to Impair Cell PolarizationFrontiers in Cell and Developmental Biology2020ISSN 2296-634X
Hasan, Wan Nuraini Wan et al.Annatto-derived tocotrienol promotes mineralization of MC3T3-E1 cells by enhancing BMP-2 protein expression via inhibiting RhoA activation and HMG-CoA reductase gene expressionDrug Design, Development and Therapy2020ISSN 1177-8881
Krueger, Irena et al.Reelin amplifies glycoprotein VI activation and alphaiib beta3 integrin outside-in signaling via PLC Gamma 2 and Rho GTPasesArteriosclerosis, Thrombosis, and Vascular Biology2020ISSN 1524-4636
Porter, Lauren et al.SUN1/2 Are Essential for RhoA/ROCK-Regulated Actomyosin Activity in Isolated Vascular Smooth Muscle CellsCells2020ISSN 2073-4409
Fox, Megan E. et al.Dendritic remodeling of D1 neurons by RhoA/Rho-kinase mediates depression-like behaviorMolecular Psychiatry2020ISSN 1476-5578
Bhosle, Vikrant K. et al.SLIT2/ROBO1-signaling inhibits macropinocytosis by opposing cortical cytoskeletal remodelingNature Communications2020ISSN 2041-1723
Salgado-Lucio, Monica L. et al.FAK regulates actin polymerization during sperm capacitation via the ERK2/GEF-H1/RhoA signaling pathwayJournal of Cell Science2020ISSN 1477-9137
Majolée, Jisca et al.CSN5 inhibition triggers inflammatory signaling and Rho/ROCK-dependent loss of endothelial integrityScientific Reports2019ISSN 2045-2322
Dias Gomes, Martim et al.Polarity signaling ensures epidermal homeostasis by coupling cellular mechanics and genomic integrityNature Communications2019ISSN 2041-1723
Li, Xu et al.A positive feedback loop of profilin-1 and RhoA/ROCK1 promotes endothelial dysfunction and oxidative stressOxidative Medicine and Cellular Longevity2018ISSN 1942-0994
Kapfhamer, Joshua et al.The Gα q/11 -provoked induction of Akr1c18 in murine luteal cells is mediated by phospholipase CMolecular and Cellular Endocrinology2018ISSN 1872-8057
Sun, Xiao Qing et al.Total flavones of Rhododendron simsii Planch flower protect isolated rat heart from ischaemia-reperfusion injury and its mechanism of UTR-RhoA-ROCK pathway inhibitionJournal of Pharmacy and Pharmacology2018ISSN 2042-7158
Santhana Kumar, Karthiga et al.TGF-β Determines the Pro-migratory Potential of bFGF Signaling in MedulloblastomaCell Reports2018ISSN 2211-1247
Tagashira, Toru et al.Afadin facilitates vascular endothelial growth factor-induced network formation and migration of vascular endothelial cells by inactivating Rho-associated kinase through ARHGAP29Arteriosclerosis, Thrombosis, and Vascular Biology2018ISSN 1524-4636
Ng, Tze Hann et al.The Rho signalling pathway mediates the pathogenicity of AHPND-causing V. parahaemolyticus in shrimpCellular Microbiology2018ISSN 1462-5822
Vautrin-Glabik, Alexia et al.IP3R3 silencing induced actin cytoskeletal reorganization through ARHGAP18/RhoA/mDia1/FAK pathway in breast cancer cell linesBiochimica et Biophysica Acta - Molecular Cell Research2018ISSN 1879-2596
Schillaci, Odessa et al.Exosomes from metastatic cancer cells transfer amoeboid phenotype to non-metastatic cells and increase endothelial permeability: Their emerging role in tumor heterogeneityScientific Reports2017ISSN 2045-2322
Kempf, Anissa et al.Control of Cell Shape, Neurite Outgrowth, and Migration by a Nogo-A/HSPG InteractionDevelopmental Cell2017ISSN 1878-1551
Yu, Yonghao et al.Hydrogen-rich medium ameliorates lipopolysaccharide-induced barrier dysfunction via rhoa-mdia1 signaling in caco-2 cellsShock2016ISSN 1540-0514
Lu, Wen Juan et al.Senescence Mediated by p16INK4a Impedes Reprogramming of Human Corneal Endothelial Cells into Neural Crest ProgenitorsScientific Reports2016ISSN 2045-2322
Rom, Slava et al.PARP inhibition in leukocytes diminishes inflammation via effects on integrins/cytoskeleton and protects the blood-brain barrierJournal of Neuroinflammation2016ISSN 1742-2094
Xiao, Bin et al.Extracellular translationally controlled tumor protein promotes colorectal cancer invasion and metastasis through Cdc42/JNK/ MMP9 signalingOncotarget2016ISSN 1949-2553
Jackson, William D. et al.Very-low and low-density lipoproteins induce neutral lipid accumulation and impair migration in monocyte subsetsScientific Reports2016ISSN 2045-2322
López-Posadas, Rocío et al.Rho-A prenylation and signaling link epithelial homeostasis to intestinal inflammationJournal of Clinical Investigation2016ISSN 1558-8238
Liu, Chunqiao et al.A secreted WNT-ligand-binding domain of FZD5 generated by a frameshift mutation causes autosomal dominant colobomaHuman Molecular Genetics2016ISSN 1460-2083
Tang, Xiao et al.HCLOCK Causes Rho-Kinase-Mediated Endothelial Dysfunction and NF-κ B-Mediated Inflammatory ResponsesOxidative Medicine and Cellular Longevity2015ISSN 1942-0994
Cascio, Graciela et al.CXCL12 Regulates through JAK1 and JAK2 Formation of Productive Immunological SynapsesThe Journal of Immunology2015ISSN 0022--1767
Manukyan, Arkadi et al.A complex of p190RhoGAP-A and anillin modulates RhoA-GTP and the cytokinetic furrow in human cellsJournal of Cell Science2015ISSN 1477-9137
Rom, Slava et al.The dual action of poly(ADP-ribose) polymerase -1 (PARP-1) inhibition in HIV-1 infection: HIV-1 ltr inhibition and diminution in Rho GTPase activityFrontiers in Microbiology2015ISSN 1664-302X
Freeman, Spencer A. et al.Toll-like receptor ligands sensitize B-cell receptor signalling by reducing actin-dependent spatial confinement of the receptorNature Communications2015ISSN 2041-1723
Rom, Slava et al.Poly(ADP-ribose) polymerase-1 inhibition in brain endothelium protects the blood-brain barrier under physiologic and neuroinflammatory conditionsJournal of Cerebral Blood Flow and Metabolism2015ISSN 1559-7016
Ochoa-Alvarez, Jhon A. et al.Antibody and lectin target podoplanin to inhibit oral squamous carcinoma cell migration and viability by distinct mechanismsOncotarget2015ISSN 1949-2553
Skrbic, Biljana et al.Lack of collagen VIII reduces fibrosis and promotes early mortality and cardiac dilatation in pressure overload in miceCardiovascular Research2015ISSN 1755-3245
Aifuwa, Ivie et al.Senescent stromal cells induce cancer cell migration via inhibition of RhoA/ROCK/myosin-based cell contractilityOncotarget2015ISSN 1949-2553
Zhu, Ying Ting et al.Knockdown of both p120 catenin and kaiso promotes expansion of human corneal endothelial monolayers via rhoa-rock-noncanonical BMP-NFκB pathwayInvestigative Ophthalmology and Visual Science2014ISSN 1552-5783
Chen, Xiaofei et al.The TMEFF2 tumor suppressor modulates integrin expression, RhoA activation and migration of prostate cancer cellsBiochimica et Biophysica Acta - Molecular Cell Research2014ISSN 1879-2596
Herr, Michael J. et al.Tetraspanin CD9 regulates cell contraction and actin arrangement via RhoA in human vascular smooth muscle cellsPLoS ONE2014ISSN 1932-6203
Biechler V., Stefanie V. et al.The impact of flow-induced forces on the morphogenesis of the outflow tractFrontiers in Physiology2014ISSN 1664-042X
Zhang, Di et al.Co-expression of delta-catenin and RhoA is significantly associated with a malignant lung cancer phenotypeInternational Journal of Clinical and Experimental Pathology2014ISSN 1936-2625
Mackay, Joanna L. et al.Simultaneous and independent tuning of RhoA and Rac1 activity with orthogonally inducible promotersIntegrative Biology (United Kingdom)2014ISSN 1757-9708
Beveridge, Ryan D. et al.The leukemia-associated Rho guanine nucleotide exchange factor LARG is required for efficient replication stress signalingCell Cycle2014ISSN 1551-4005
Ahn, Bum Ju et al.Ninjurin1 enhances the basal motility and transendothelial migration of immune cells by inducing protrusive membrane dynamicsJournal of Biological Chemistry2014ISSN 1083-351X
Papke, Christina L. et al.Smooth muscle hyperplasia due to loss of smooth muscle α-actin is driven by activation of focal adhesion kinase, altered p53 localization and increased levels of platelet-derived growth factor receptor-βHuman Molecular Genetics2013ISSN 0964-6906
Kanazawa, Yasushi et al.The Rho-kinase inhibitor fasudil restores normal motor nerve conduction velocity in diabetic rats by assuring the proper localization of adhesion-related molecules in myelinating Schwann cellsExperimental neurology2013ISSN 1090--2430
Tan, Hong et al.Fluid flow forces and rhoA regulate fibrous development of the atrioventricular valvesDevelopmental Biology2013ISSN 1095-564X
DiScipio, Richard G. et al.Complement C3a signaling mediates production of angiogenic factors in mesenchymal stem cellsJournal of Biomedical Science and Engineering2013ISSN 1937--6871
Kalia, Manjula et al.Japanese Encephalitis Virus Infects Neuronal Cells through a Clathrin-Independent Endocytic MechanismJournal of Virology2013ISSN 0022--538X
Yang, Jian et al.Inhibition of farnesyl pyrophosphate synthase attenuates angiotensin II-induced cardiac hypertrophy and fibrosis in vivoInternational Journal of Biochemistry and Cell Biology2013ISSN 1357-2725
Rom, Slava et al.Selective activation of cannabinoid receptor 2 in leukocytes suppresses their engagement of the brain endothelium and protects the blood-brain barrierAmerican Journal of Pathology2013ISSN 0002-9440
Dubash, Adi D. et al.The GEF Bcr activates RhoA/MAL signaling to promote keratinocyte differentiation via desmoglein-1Journal of Cell Biology2013ISSN 0021-9525
Elali, Ayman et al.Liver X receptor activation enhances blood-brain barrier integrity in the ischemic brain and increases the abundance of ATP-binding cassette transporters ABCB1 and ABCC1 on brain capillary cellsBrain Pathology2012ISSN 1015-6305
Zhu, Ying Ting et al.Nuclear p120 catenin unlocks mitotic block of contactinhibited human corneal endothelial monolayers without disrupting adherent junctionsJournal of Cell Science2012ISSN 0021-9533
Greco, Carolina M. et al.Chemotactic effect of prorenin on human aortic smooth muscle cells: a novel function of the (pro)renin receptorCardiovascular Research2012ISSN 0008--6363
Dhaliwal, Anandika et al.Cellular Cytoskeleton Dynamics Modulates Non-Viral Gene Delivery through RhoGTPases2012PMID 22509380
Ridgway, Lon D. et al.Heparanase-induced GEF-H1 signaling regulates the cytoskeletal dynamics of brain metastatic breast cancer cellsMolecular Cancer Research2012ISSN 1541-7786
Chen, Si Meng et al.Inhibition of tumor cell growth, proliferation and migration by X-387, a novel active-site inhibitor of mTORBiochemical Pharmacology2012ISSN 0006-2952
Chen, Guang et al.Inhibition of chemokine (CXC motif) ligand 12/chemokine (CXC motif) receptor 4 axis (CXCL12/CXCR4)-mediated cell migration by targeting mammalian target of rapamycin (mTOR) pathway in human gastric carcinoma cells (Journal of Biological Chemistry (2012) 2Journal of Biological Chemistry2012ISSN 0021-9258
Howe, Grant A. et al.RhoB controls endothelial cell morphogenesis in part via negative regulation of RhoAVascular Cell2012ISSN 2045--824X
Yang, Seungwon et al.The RhoA-ROCK-PTEN pathway as a molecular switch for anchorage dependent cell behaviorBiomaterials2012ISSN 1878--5905
Garrido‐Gómez, Tamara et al.Annexin A2 is critical for embryo adhesiveness to the human endometrium by RhoA activation through F‐actin regulationThe FASEB Journal2012ISSN 0892--6638
Zhou, Zhigang et al.HSV-mediated gene transfer of C3 transferase inhibits Rho to promote axonal regenerationExperimental Neurology2012ISSN 0014--4886
McCoy, Kelly L. et al.Protease-activated receptor 1 (PAR1) coupling to G(q/11) but not to G(i/o) or G(12/13) is mediated by discrete amino acids within the receptor second intracellular loopCellular signalling2012ISSN 1873--3913
Ramseyer, Vanesa D. et al.Tumor necrosis factor α decreases nitric oxide synthase type 3 expression primarily via Rho/Rho kinase in the thick ascending limbHypertension (Dallas, Tex. : 1979)2012ISSN 1524--4563
Takefuji, Mikito et al.G13-mediated signaling pathway is required for pressure overload-induced cardiac remodeling and heart failureCirculation2012ISSN 0009-7322
Jin, Wanzhu et al.Increased SRF transcriptional activity in human and mouse skeletal muscle is a signature of insulin resistanceJournal of Clinical Investigation2011ISSN 0021-9738
Aguilar, Hector N. et al.Phos-tag-based analysis of myosin regulatory light chain phosphorylation in human uterine myocytesPLoS ONE2011ISSN 1932-6203
Mammoto, Tadanori et al.Mechanochemical Control of Mesenchymal Condensation and Embryonic Tooth Organ FormationDevelopmental Cell2011ISSN 1534-5807
Ganguly, Riya et al.Adiponectin Increases LPL Activity via RhoA/ROCK-Mediated Actin Remodelling in Adult Rat CardiomyocytesEndocrinology2011ISSN 0013--7227
Musso, Alessandra et al.Relevance of the mevalonate biosynthetic pathway in the regulation of bone marrow mesenchymal stromal cell-mediated effects on T-cell proliferation and B-cell survivalHaematologica2011ISSN 1592--8721
Yang, Enyue et al.Fluoride induces vascular contraction through activation of RhoA/Rho kinase pathway in isolated rat aortasEnvironmental toxicology and pharmacology2010ISSN 1872--7077
Ridgway, Lon D. et al.Modulation of GEF-H1 Induced Signaling by Heparanase in Brain Metastatic Melanoma CellsJournal of cellular biochemistry2010ISSN 0730-2312
Nini, Lylia et al.Accurate and reproducible measurements of RhoA activation in small samples of primary cellsAnalytical biochemistry2010ISSN 1096--0309
Rapier, Rebecca et al.The extracellular matrix microtopography drives critical changes in cellular motility and Rho A activity in colon cancer cellsCancer Cell International2010ISSN 1475-2867
Lichtenstein, Mathieu P. et al.Secretase-independent and RhoGTPase/PAK/ERK-dependent regulation of cytoskeleton dynamics in astrocytes by NSAIDs and derivativesJournal of Alzheimer's disease : JAD2010ISSN 1875--8908
Romero, Ana M. et al.Chronic ethanol exposure alters the levels, assembly, and cellular organization of the actin cytoskeleton and microtubules in hippocampal neurons in primary cultureToxicological Sciences2010ISSN 1096-6080
Hammar, Eva et al.Role of the Rho-ROCK (Rho-Associated Kinase) Signaling Pathway in the Regulation of Pancreatic β-Cell FunctionEndocrinology2009ISSN 0013--7227
Liu, Yang et al.Ablation of p120-catenin enhances invasion and metastasis of human lung cancer cellsCancer Science2009ISSN 1347-9032
Seifert, Jennifer L. et al.Differential activation of Rac1 and RhoA in neuroblastoma cell fractionsNeuroscience letters2009ISSN 0304--3940
Chastre, Eric et al.TRIP6, a novel molecular partner of the MAGI-1 scaffolding molecule, promotes invasivenessThe FASEB Journal2009ISSN 1530--6860
Mercer, Jason et al.Vaccinia virus uses macropinocytosis and apoptotic mimicry to enter host cellsScience (New York, N.Y.)2008ISSN 1095--9203
Gien, Jason et al.Chronic intrauterine pulmonary hypertension increases endothelial cell Rho kinase activity and impairs angiogenesis in vitroAmerican Journal of Physiology - Lung Cellular and Molecular Physiology2008ISSN 1040-0605
Ramirez, Servio H. et al.Activation of Peroxisome Proliferator-Activated Receptor γ (PPARγ) Suppresses Rho GTPases in Human Brain Microvascular Endothelial Cells and Inhibits Adhesion and Transendothelial Migration of HIV-1 Infected MonocytesThe Journal of Immunology2008ISSN 0022--1767
Korobova, Farida et al.Arp2/3 complex is important for filopodia formation, growth cone motility, and neuritogenesis in neuronal cellsMolecular biology of the cell2008ISSN 1939--4586
Moore, Simon W. et al.Rho inhibition recruits DCC to the neuronal plasma membrane and enhances axon chemoattraction to netrin 1Development2008ISSN 0950--1991
Kinoshita, Nagatoki et al.Apical Accumulation of Rho in the Neural Plate Is Important for Neural Plate Cell Shape Change and Neural Tube FormationMolecular Biology of the Cell2008ISSN 1059-1524
Sequeira, Linda et al.Rho GTPases in PC-3 prostate cancer cell morphology, invasion and tumor cell diapedesisClinical & experimental metastasis2008ISSN 0262--0898
Schreibelt, Gerty et al.Reactive oxygen species alter brain endothelial tight junction dynamics via RhoA, PI3 kinase, and PKB signalingFASEB journal : official publication of the Federation of American Societies for Experimental Biology2007ISSN 1530--6860
Keely, Patricia J. et al.Investigating integrin regulation and signaling events in three-dimensional systemsMethods in enzymology2007ISSN 0076--6879
Tanaka, Shigeru et al.Neural expression of G protein-coupled receptors GPR3, GPR6, and GPR12 up-regulates cyclic AMP levels and promotes neurite outgrowthThe Journal of biological chemistry2007ISSN 0021--9258
Rupp, Paul A. et al.A role for RhoA in the two-phase migratory pattern of post-otic neural crest cellsDevelopmental Biology2007ISSN 0012-1606
Bradley, William D. et al.Integrin Signaling through Arg Activates p190RhoGAP by Promoting Its Binding to p120RasGAP and Recruitment to the MembraneMolecular Biology of the Cell2006ISSN 1059-1524

Question 1: Which is the best Small G-protein activation assay for my tissue? 

Answer 1: Cytoskeleton, Inc. offers a large number of G-LISA and traditional pull-down activation assays to study the biology and biochemistry of small G-proteins.  Our G-LISA activation assays provide an improved method of measuring the activity of small G-proteins utilizing a simple and quick protocol in 96-well format to provide extremely accurate results. To complement our G-LISA line of activation assays, we also offer the most efficiently designed and complete traditional pull-down activation assays available.

 

Based on enzyme-linked immunosorbent assay (ELISA) technology, our G-LISA activation assays utilize a 96 well format with 12 x 8 well strips that provide the flexibility to run 1 to 96 wells (each well is a condition/treatment) at one time.  This flexibility is especially important if any concentration, dose or time course analyses will be performed.  Pull-downs are limited to the number of wells in each gel (usually 10-15) that will be run.  The G-LISA assays are also more accurate and quantitative than pull-downs with greater sensitivity while using less material per well.  The isotype specificity depends on the antibodies being used to capture and visualize the small G-proteins.  We have designed both types of activation assays to specifically target RhoA and Rac1.  Each of these assays can be easily modified to study RhoB and RhoC or Rac2 and Rac3, respectively.  We also offer activation assays for Cdc42, Ras, and RalA.  For a quick “look and see” experiment with a few treatment conditions (one drug concentration and time point), pull-downs are convenient.  Pull-downs also utilize methods that are very familiar to most labs and so offer a convenient entry point for small G-protein activation assays.  Besides the standard sized pull-down kits (50-80 assays, depending on protein), we also offer a starter-sized kit (20 assays) for RhoA, Rac1, Cdc42, and Ras.  To screen for multiple small G-proteins, we recommend the Combo starter pull-down kit which provides 10 assays each for RhoA, Rac1, and Cdc42.  

For an interactive guide to help choose the activation assay that is right for you, click here.

Question 2: Are there any citations for the G-LISA series?

Answer 2:Yes, the G-LISA activation assays are well-cited in the scientific literature.  Please see the “Citations” tab above for links to some current citations for our RhoA, Rac and Cdc42 G-LISA activation assays.

Select from the categories below:

  1. RhoA / Rac1 / Cdc42 Activation Assay Combo Biochem Kit (bead pull-down format) - 3 x 10 assays BK030
    RhoA / Rac1 / Cdc42 Activation Assay Combo Biochem Kit (bead pull-down format) - 3 x 10 assays
    Learn More
  2. Arf1 G-LISA Activation Assay Kit (Colorimetric Based) - 96 assays BK132

    G-LISA Arf1 Activation Assay Biochem Kit (Colorimetric Based)

    Learn More
  3. Arf1 Pull-down Activation Assay Biochem Kit (bead pull-down format) - 20 Assays BK032-S
    Arf1 Activation Assay Biochem Kit (bead pull-down format) - 20 Assays
    Learn More
  4. Arf6 Pull-down Activation Assay Biochem Kit (bead pull-down format) - 20 Assays BK033-S
    Arf6 Activation Assay Biochem Kit (bead pull-down format) - 20 Assays
    Learn More
  5. Cdc42 G-LISA Activation Assay (Colorimetric format) - 24 assays BK127-S

    G-LISA Cdc42 Activation Assay Biochem Kit (Colorimetric format)

    Learn More
  6. Cdc42 G-LISA Activation Assay (Colorimetric format) - 96 assays BK127

    G-LISA Cdc42 Activation Assay Biochem Kit (Colorimetric format)

    Learn More
  7. Cdc42 Pull-down Activation Assay Biochem Kit (bead pull-down format) - 20 Assays BK034-S
    Cdc42 Activation Assay Biochem Kit (bead pull-down format) - 20 Assays
    Learn More
  8. Cdc42 Pull-down Activation Assay Biochem Kit (bead pull-down format) - 50 Assays BK034
    Cdc42 Activation Assay Biochem Kit (bead pull down format) - 50 Assays
    Learn More
  9. Rac1 G-LISA Activation Assay (Luminescence format) - 96 assays BK126
    G-LISA Rac 1 Activation Assay Biochem Kit (luminescence format)
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  10. Rac1 Pull-down Activation Assay Biochem Kit (bead pull-down format) - 20 Assays BK035-S
    Rac1 Activation Assay Biochem Kit (bead pull-down format) - 20 Assays
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  11. Rac1 Pull-down Activation Assay Biochem Kit (bead pull-down format) - 50 Assays BK035
    Rac1 Activation Assay Biochem Kit (bead pull-down format) - 50 Assays
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  12. Rac1,2,3 G-LISA Activation Assay (Colorimetric format) 96 assays BK125
    G-LISA Rac 1,2,3 Activation Assay Biochem Kit (colorimetric format)
    Learn More
  13. Ras G-LISA Activation Assay Kit (Colorimetric Based) - 96 assays BK131
    G-LISA Ras Activation Assay Biochem Kit (Colorimetric format) Learn More
  14. Ras Pull-down Activation Assay Biochem Kit (bead pull-down format) - 20 Assays BK008-S
    Ras Activation Assay Biochem Kit (bead pull-down format) - 20 Assays
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  15. Ras Pull-down Activation Assay Biochem Kit (bead pull-down format) - 50 Assays BK008
    Ras Activation Assay Biochem Kit (bead pull-down format) - 50 Assays
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  16. RhoA G-LISA Activation Assay (Luminescence format) 96 assays BK121
    G-LISA RhoA Activation Assay Biochem Kit (luminescence format)
    Learn More
  17. RhoA G-LISA Activation Assay Kit (Colorimetric format) 24 assays BK124-S

    G-LISA RhoA Activation Assay Biochem Kit (Colorimetric format)

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  18. RhoA G-LISA Activation Assay Kit (Colorimetric format) 96 assays BK124
    G-LISA RhoA Activation Assay Biochem Kit (colorimetric format)
    Learn More
  19. RhoA Pull-down Activation Assay Biochem Kit (bead pull-down format) - 20 Assays BK036-S
    RhoA Activation Assay Biochem Kit (bead pull-down format) - 20 Assays
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  20. RhoA Pull-down Activation Assay Biochem Kit (bead pull-down format) - 80 Assays BK036
    RhoA Activation Assay Biochem Kit (bead pull-down format) - 80 Assays
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