Small G-protein Activation Assays
Small G-protein activation assays are essential tools for investigating cellular signaling pathways.
Cyto provides a broad selection of assays, including Rac1, RhoA, Ras, Cdc42, Arf1, and Arf6. These activation assays are available in two formats:
G-LISA® Assay
This format uses a 96-well strip-based ELISA approach, where an effector protein immobilized in each well selectively captures the GTPase's active (GTP-bound) form directly from cell lysates. Detection is achieved through a specific antibody-based system.
Key Benefits:- Quantitative and reproducible
- High sensitivity: requires only 10–50 µg of lysate
- Rapid results: under 3 hours
- Flexible strip-well format: use only the number of wells needed
Traditional Pull-down Assay
This method uses agarose beads conjugated to an effector domain to isolate active GTPases from cell lysates. The pulled-down proteins are then analyzed via SDS-PAGE and western blot using GTPase-specific antibodies.
Key Benefits:- Widely used and well-established
- Allows downstream analysis of interacting proteins
Both formats are user-friendly and comprehensive, with G-LISA® standing out for its speed, sensitivity, and quantitative nature, making it particularly useful for high-throughput or in systems with limited lysate, such as primary cell cultures.
Small G-protein activation assays are essential tools for investigating cellular signaling pathways.
Cyto provides a broad selection of assays, including Rac1, RhoA, Ras, Cdc42, Arf1, and Arf6. These activation assays are available in two formats:
G-LISA® Assay
This format uses a 96-well strip-based ELISA approach, where an effector protein immobilized in each well selectively captures the GTPase's active (GTP-bound) form directly from cell lysates. Detection is achieved through a specific antibody-based system.
Key Benefits:- Quantitative and reproducible
- High sensitivity: requires only 10–50 µg of lysate
- Rapid results: under 3 hours
- Flexible strip-well format: use only the number of wells needed
Traditional Pull-down Assay
This method uses agarose beads conjugated to an effector domain to isolate active GTPases from cell lysates. The pulled-down proteins are then analyzed via SDS-PAGE and western blot using GTPase-specific antibodies.
Key Benefits:- Widely used and well-established
- Allows downstream analysis of interacting proteins
Both formats are user-friendly and comprehensive, with G-LISA® standing out for its speed, sensitivity, and quantitative nature, making it particularly useful for high-throughput or in systems with limited lysate, such as primary cell cultures.
