Choosing an activation assay

 

Step 1: Initial Considerations (Equipment, Starting Material and Cost)

The following should be considered when choosing the right activation assay for your needs.  Please continue to step 2 below  once you have reviewed these specifications.

Assay Factor

 

Assay Considerations

 

Recommendations

Equipment    Traditional Pull-down
  • SDS-PAGE apparatus
  • Western blot apparatus

 

  Consider the equipment requirements prior to deciding upon an assay format.
  G-LISA
  • Orbital Plate shaker
  • Spectrophotometer (set to 490 nm)
  • Multichannel pipette (recommended if handling high number of samples)
 

Starting material required per assay

 

 
Traditional Pull-down
  • 300-2000 µg per assay

 

  If starting material is limited, e.g. primary cells, the G-LISA assay is required.
  G-LISA
  • 5-50 µg per assay
 
Cost   
Traditional Pull-down
  • Combo kit (10 assays each RhoA/Rac1/Cdc42)
  • Starter kit (20 assays)
  • Regular kit (50-80 assays depending on target protein)

 

 

Cost per assay:  The G-LISA offers the best value per assay, particularly if you anticipate performing >50 assays.

 

Cost per kit:  The traditional pull-down assay starter kits generally provide the lowest cost per kit.  NOTE: Consider the combo kit as a cost effective choice if you require a quick assessment of the role of RhoA/Rac1/Cdc42 in your system.

  G-LISA
  • Available in 96 well format.  Each well can be broken off and used individually.
 

 

Step 2: Choose the number of assays required.


1 - 3

4 - 20

21 - 1000