G-LISA Bundle (BK135 = BK124-S + BK127-S + BK128-S) (colorimetric)

Highlights

• The RhoA / Rac1 / Cdc42 G-LISA Activation Assay Bundle is a cost effective choice when your experimental observations suggest that one or more Rho family proteins are activated.
• Provides 24 assays each for detection of activated RhoA, Rac1, and Cdc42.
• Includes an extensive array of reagents (see datasheet for individual kit contents).
• Most cost effective and precise method to obtain a Rho family activation profile.

Introduction
The G-LISA activation assays are ELISA-based activation assays with which you can measure RhoA, Rac1, and Cdc42 activity in as little as 10 ug of cell or tissue lysates in less than 3 hours. These assays are very sensitive and have excellent accuracy between duplicate samples. For a more detailed introduction on G-LISA assays and a listing of other available G-LISA kits, see our main G-LISA page.

Kit contents
Each kit contains sufficient reagents to perform 24 activation assays. Larger 96-well assays are also available (see related products). Since the GTP affinity wells are supplied as 12 x 8 well strips and the strips can be broken into smaller pieces, each kit can be used for anywhere from one to multiple assays. For a list of components, please view each of the kit manuals below.

1. RhoA G-LISA Activation Assay Manual (view pdf here)
2. Rac1 G-LISA Activation Assay Manual (view pdf here)
3. Cdc42 G-LISA Activation Assay Manual (view pdf here)

Equipment needed

1. 96-well plate spectrophotometer capable of reading 490 nm wavelength
2. Orbital microplate shaker capable of at least 200 rpm shaking (400 rpm is optimal)
3. Multichannel or multidispensing pipettor (optional)

Example results

Figure 1. RhoA activation by calpeptin measured by G-LISA kit BK124. Swiss 3T3 (mouse) cells were serum starved for 24 h and treated with calpeptin (Cal; 0.1 mg/ml for 30 min) or DMSO carrier only (SS). 10 ug of cell lysates were subjected to the G-LISA assay. Absorbance was read at 490 nm.

Figure 2. Cdc42 activation by EGF measured by the Cdc42 G-LISA Activation Assay. Swiss 3T3 cells were serum starved (SS) for 16 h at 1% serum and 8 h with 0% serum and treated with EGF (100 ng/ml for 2 min). Cell lysates (8, 17, 35 ug) were subjected to the G-LISA assay. Data was read at 490 nm. Numbers on top the yellow columns indicate the fold increase in signal caused by EGF activation, you will notice the ratio remains the same with different protein loadings indicating good linearity with different protein loadings. 500 ug of the same lysates were subjected to the traditional PAK pull-down assay (Cat.# BK034) with similar results.

Associated Products:

Total RhoA ELISA (Cat. # BK150)

Anti-RhoA monoclonal antibody (Cat.# ARH03)

Rho Inhibitor I (Cat.# CT04)

Rho Activator II (Cat.# CN03)

Rho Pathway Inhibitor I - Rho Kinase (ROCK) inhibitor Y-27632 (Cat. # CN06)