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G lisa cdc42 activation assay biochem kit

Manufactured by Cytoskeleton
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The G-LISA Cdc42 Activation Assay Biochem Kit is a laboratory equipment product designed to detect and quantify the activation of the Cdc42 protein, a member of the Rho family of small GTPases, in biological samples. The kit utilizes a Cdc42-GTP-binding protein linked to the wells of a 96-well plate to capture and measure the active, GTP-bound form of Cdc42. The amount of active Cdc42 is then detected colorimetrically, providing a quantitative assessment of Cdc42 activation.

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Lab products found in correlation

Rabbit anti phospho jnk, by Abcam (1 mentions) Mouse anti gapdh, by Zhongshan Golden Bridge Biotechnology (1 mentions) Rabbit anti jnk, by Abcam (1 mentions) Mouse anti tubulin, by Proteintech (1 mentions) Rabbit anti igg, by Santa Cruz Biotechnology (1 mentions) G lisa rhoa activation assay biochem kit, by Cytoskeleton (1 mentions) G lisa rac1 activation assay biochem kit, by Cytoskeleton (1 mentions)

Close spelling variants of this product

Cdc42 G-LISA Activation Assay Kit Cdc42 Activation Assay Biochem Kit G-LISA activation assay kit G-LISA Activation Assay Cdc42 G-Lisa G-LISA assay G-LISA kit G-LISA Cdc42

7 protocols using g lisa cdc42 activation assay biochem kit

1

Assessing Cdc42 Activation in 3D Cysts

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Endogenous Cdc42 activity was determined by GST-PBD pull-down assay. Briefly, the lysates were incubated at 4 °C for 1 h with GST-PBD beads and the amounts of total and active Cdc42 were detected by western blot. G-LISA Cdc42 Activation Assay Biochem Kit (Cytoskeleton, BK127) were used to measure levels of activated Cdc42 in 3D cysts according to the manufacturer's instructions.
Tien S.C., Lee H.H., Yang Y.C., Lin M.H., Chen Y.J, & Chang Z.F. (2016). The Shp2-induced epithelial disorganization defect is reversed by HDAC6 inhibition independent of Cdc42. Nature Communications, 7, 10420.
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2

HUVEC activation by S. aureus

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HUVEC were serum starved in base media containing DMSO or ML 141 (18-20 h). S. aureus cultures were washed as described above, incubated with FBS (15 min, RT) as the source of fibronectin, washed extensively, resuspended in saline, and incubated with HUVEC (MOI 1440, 1 h, 5% CO2, 37°C). Total protein concentration of lysates was determined, samples diluted to 0.125 mg/ml in lysis buffer containing protease inhibitors, and GTP-bound CDC42 detected using the G-LISA Cdc42 Activation Assay Biochem Kit (Cytoskeleton, Denver, CO). Of note, more concentrated lysates (0.25 mg/ml) appeared to saturate the assay.
Cordero D., Fullenkamp C.R., Pelly R.R., Reed K.M., Caffo L.M., Zahrt A.N., Newman M., Komanapalli S., Niemeier E.M., Bishop D.L., Bruns H.A., Haynes M.K., Sklar L.A., Sammelson R.E, & McDowell S.A. (2014). Small Molecule Inhibitors Limit Endothelial Cell Invasion by Staphylococcus aureus. Current pharmaceutical biotechnology, 15(8), 727-737.
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3

Investigating Cytoskeletal Regulation in Muscle

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The primary antibodies used in the study included rabbit anti-Profilin1 (1:1000, Abcam, ab50667), rabbit anti-Cdc42 (1:1000, Proteintech, Rosemont, IL, USA), rabbit anti-PAK (1:1000, Proteintech, Rosemont, IL, USA), rabbit anti-Phospho PAK (1:1500, Proteintech, IL, USA), rabbit anti-IgG (1:50, Santa Cruz, Dallas, TX, USA), rabbit anti-JNK (1:1000, Abcam, ab179461, London, UK), rabbit anti-Phospho JNK(1:1000, Abcam, ab124956, London, UK), mouse anti-MyHC (1:100, DSHB, CO, USA), mouse anti-MyoG (1:100, DSHB, Iowa, IA, USA), mouse anti-GAPDH (1:1000, Zhongshan, Golden Bridge Bio-technology, Beijing, China), and mouse anti-tubulin (1:3000, Proteintech, Rosemont, IL, USA). The secondary antibodies included horseradish peroxidase goat anti-mouse/rabbit IgG (1:10000, Zhongshan, Golden Bridge Bio-technology, Beijing, China), PAK inhibitors (Selleck, PF-3758309), and JNK inhibitors (GLPBIO, SP600125). A G-LISA® Cdc42 Activation Assay Biochem Kit™ (cytoskeleton, Cat. # BK127-S) was used.
Zi J., Xu J., Luo J., Yang X., Zhen Z., Li X., Hu D., Guo Y., Guo H., Ding X, & Zhang L. (2022). PFN1 Inhibits Myogenesis of Bovine Myoblast Cells via Cdc42-PAK/JNK. Cells, 11(20), 3188.
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4

Quantifying Active Cdc42 Levels

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Active Cdc42 was measured by G-LISA Cdc42 Activation Assay Biochem kit (colorimetric assay, Cytoskeleton, Denver, CO, USA) following the manufacturer's instruction. And the signal was measured at 490 nm with a microplate reader (MRX, Dynatech Laboratories, Chantilly, VA, USA). Total Cdc42 protein expression of each group was measured by WB, as described above. Cell lysate from each group containing the same amount of protein was measured in both Cdc42 GTPase activation assay and WB. Results were expressed as fold activity of stimulated in relation to non-stimulated controls normalized to total Cdc42 protein content.
Huang Z., Zhang L., Chen Y., Zhang H., Zhang Q., Li R., Ma J., Li Z., Yu C., Lai Y., Lin T., Zhao X., Zhang B., Ye Z., Liu S., Wang W., Liang X., Liao R, & Shi W. (2016). Cdc42 deficiency induces podocyte apoptosis by inhibiting the Nwasp/stress fibers/YAP pathway. Cell Death & Disease, 7(3), e2142-.
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5

Quantifying CDC42 and RAC1 Activity

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CDC42-GTP (Active-CDC42) and Active-RAC1 levels in organoids before and after E2 and DBZ treatment were determined using RAC1/CDC42 Activation Magnetic Beads Pull-down Assay Kits from Merck Millipore following the instructions. In brief, 100 μg of clarified total lysates were incubated with PAK-1 PBD magnetic beads in microfuge tube at 4 °C for 1 h, and the amount of total and active CDC42 or RAC1 were detected by western blot. G-LISA Cdc42 Activation Assay Biochem Kit (Cytoskeleton, BK127) was used to measure levels of activated CDC42 in organoids treated with CDC42 activity inhibitors was determined using according to the manufacturer’s instructions.
Jiang R., Tang X., Pan J., Li G., Yang N., Tang Y., Bi S., Cai H., Chen Q., Chen D., Wang H, & Kong S. (2022). CDC42 governs normal oviduct multiciliogenesis through activating AKT to ensure timely embryo transport. Cell Death & Disease, 13(9), 757.
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6

Rac1, RhoA, and Cdc42 Activation Assay

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KDR cells at 50%confluence were incubated for 24 h in FCS-free DMEM to starve the cells, and then treated with 10% FCS for 30 min. Cell lysates were collected and levels of activated GTP-bound Rac1, RhoA and Cdc42 were determined using the G-LISA Rac1 Activation Assay Biochem Kit (BK126, Cytoskeleton Inc., CO, USA), G-LISA RhoA Activation Assay Biochem Kit (BK121, Cytoskeleton Inc.) and G-LISA Cdc42 Activation Assay Biochem Kit (BK127, Cytoskeleton Inc.), respectively.
Yan Y., Tsukamoto O., Nakano A., Kato H., Kioka H., Ito N., Higo S., Yamazaki S., Shintani Y., Matsuoka K., Liao Y., Asanuma H., Asakura M., Takafuji K., Minamino T., Asano Y., Kitakaze M, & Takashima S. (2015). Augmented AMPK activity inhibits cell migration by phosphorylating the novel substrate Pdlim5. Nature Communications, 6, 6137.
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7

Cdc42 Activation in MDA-MB-231 Cells

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To detect GTP-active bound Cdc42, MDA-MB-231 transfected with CT or DDR1 #1 siRNA were cultured for 2 h on type I collagen fibrils or overnight on plastic. 50 µg of protein was subjected using the G-LISA Cdc42 Activation Assay Biochem kit (Cytoskeleton, Inc.) according to the manufacturer’s instructions.
Juin A., Di Martino J., Leitinger B., Henriet E., Gary A.S., Paysan L., Bomo J., Baffet G., Gauthier-Rouvière C., Rosenbaum J., Moreau V, & Saltel F. (2014). Discoidin domain receptor 1 controls linear invadosome formation via a Cdc42–Tuba pathway. The Journal of Cell Biology, 207(4), 517-533.
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