The G-LISA for 3D media cultures

In the research communities of neuroscience and cancer research, it is increasingly recognized that three-dimensional (3D) cell cultures (e.g. organoids and gelatinous media) offer significant advantages over two-dimensional (2D) cell cultures in preclinical studies and drug discovery studies. 3D cultures recap the properties of the environment found in vivo with extracellular matrix (ECM) in more detail (see figure).

One of the main obstacles in 3D and 2D cultures is the reliable and sensitive quantification of the GTPase activation levels of Ras and Rho in various cell culture models for cancer. ELISA-based G-LISA activation assays offer several significant advantages over conventional pulldown activation assays.

Advantages of G-LISAs over pull-downs for 3D culture activation assays
less labor intensive.
only require 50,000 to 200,000 cells per sample, while pulldowns require 10 to 20 million cells.
enable multi-point studies, while pulldowns are limited to a few points.
Applying small G-protein G-LISA activation assays to 3D culture systems provides flexibility across multiple parameters, including the types of 3D matrices and cells used and the small GTPases analyzed.

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