pulldownglisaApplications for pull-downs


Measure K-Ras, N-Ras, H-Ras, RhoC, Rac3 isoforms

  • Active levels of multiple Ras- and Rho-subfamily GTPase isoforms can be detected

Complement G-LISA findings

  • Use two different, validated techniques to quantitate active GTPase level.


Applications for G-LISAs


Swiss 3T3 fibroblasts were treated with 1 μg/ml CN04 for the indicated times and cell lysates were subjected to G-LISA activation assays for RhoA (Cat.# BK124), Rac1 (Cat.# BK128), and Cdc42 (Cat.# BK127).

Measure active GTPases in 3-D cell cultures

Measure active GTPases in primary cell lysates

  • Measure active GTPases in as little as 5 µg of total cell extract.

Increase accuracy of active GTPase quantitations

  • Quantitative numerical output in OD490nm units.

Measure active GTPases in tissue lysates

  • Use as little as 1 mm3 of tissue for extraction.


Cited in hundreds of peer- reviewed articles

3-D Cell Culture and G-LISAs: 


Ponik S.M. et al. 2013. RhoA is down-regulated at cell-cell contacts via p190RhoGAP-B in response to tensional homeostasis. Mol. Biol. Cell. 24, 1688-1699. 


Petroll W.M. et al. 2008. Dynamic assessment of fibroblast mechanical activity during Rac-induced cell spreading in 3-D culture. J. Cell Physiol. 217, 162-171. 


Keely P.J. et al. 2007. Investigating integrin regulation and signaling events in three-dimensional systems. Methods Enzymol. 426, 27-45. 



Primary Cells and G-LISAs: 


Valdez C.M. et al. 2016. The Rac-GAP alpha2-chimaerin regulates hippocampal dendrite and spine morphogenesis.  Mol. Cell. Neurosci. 75, 14-26. 

Nini L. and Dagino L. 2010. Accurate and reproducible measurements of RhoA activation in small samples of primary cells. Anal. Biochem. 398, 135-137. 

Flaiz C. et al. 2009. PAK kinase regulates Rac GTPase and is a potential target in human schwannomas. Exp. Neurol. 218, 137-144. 


Increased Accuracy and G-LISAs:


Oliver A.W. et al. 2011. The HPV16 E6 binding protein Tip-1 interacts with ARHGEF16, which activates Cdc42. Br. J. Cancer. 104, 324 - 331. 


GTPase Isoforms and Pull-downs:


Tabibian J.H. et al. 2014. Cholangiocyte senescence by way of N-Ras activation is a characteristic of primary sclerosing cholangitis. Hepatology. 59, 2263-2275. 

Kusuhara S. et al. 2012. Arhgef15 promotes retinal angiogenesis by mediating VEGF-induced Cdc42 activation and potentiating RhoJ inactivation in endothe­lial cells. PLoS ONE. 9:e45858. 


Pull-downs Complementing G-LISAs:


Tanaka T. et al. 2010. Monocyte chemoattractant protein-1/CC chemokine ligand 2 enhances apoptotic cell removal by macrophages through Rac1 activation. Biochem. Biophys. Res. Commun. 399, 677-682. 


Schlegel N. and Waschke J. 2009. VASP is involved in cAMP-mediated Rac 1 activation in microvascular endothelial cells. Am. J. Physiol. Cell Physiol. 296, C453-C462. 


Lai F.P.L. et al. 2009. Cortactin Promotes Migration and Platelet-derived Growth Factor-induced Actin Reorganization by Signaling to Rho-GTPases. Mol. Biol. Cell. 20, 3209-3223. 


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