Ras G-LISA Activation Assay Kit (colorimetric)

AT

With the new Ras G-LISA kit you can now measure Ras activation from cell and tissue samples in less than 3 h. G-LISA requires only 1-5% of the material needed for a conventional pull-down assay. You will also be able to handle large sample numbers and generate quantitative results. For a more detailed introduction on G-LISA assays and a listing of other available G-LISA kits, see our main G-LISA page.
The Ras G-LISA kit contains a Ras GTP-binding protein linked to the wells of a 96 well plate. Active, GTP-bound Ras in cell/tissue lysates will bind to the wells while inactive GDP-bound Ras is removed during washing steps. The bound active Ras is detected with a Ras specific antibody. The degree of Ras activation is determined by comparing readings from activated cell lysates versus non-activated cell lysates. Inactivation of Ras is generally achieved in tissue culture by a serum starvation step (see kit datasheet for more information).

1-2 Weeks

950

96-well plate spectrophotometer capable of reading 490 nm wavelength
Multichannel or multidispensing pipetto
Orbital microplate shaker capable of at least 200 rpm shaking (400 rpm is optimal)

Ras activation by EGF measured by G-LISA. HeLa cells were serum starved (SS) for 24 h and treated with EGF (100 ng/ml for 2 min). 25, 12.5, 5, 1.25 ug of cell lysates were subjected to the G-LISA assay. Absorbance was read at 490 nm. Data are background subtracted