Traditional pull-down assays use methods familiar to most labs and therefore provide a familiar point of entry.
The G-LISA assay is simple to use and has many advantages that should be considered (see below for comparisons).
Consider the equipment requirements prior to deciding upon an assay format.
Starting material required per assay
300 - 2000 µg per assay
If starting material is limited, e.g. primary cells, the G-LISA assay is required.
5 – 50 µg per assay
Cost issues to consider:
Cost per assay: The G-LISA is the best value per assay.
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.
Available in 96 well format. Each well can be broken off and used individually.
Centrifugation steps and western blot analysis makes this assay most suitable for low sample numbers (≤10).
<10 samples: either assay
>10 samples: G-LISA recommended
NOTE: Dose responses and time courses are often recommended and will greatly increase the number of samples being analyzed.
Amenable to high sample numbers (>10) or dose response/time-course analyses. Wells can be detached and used individually.
Quantification of results
Western blot quantification has a very narrow linear range. Also, multiple manipulations of the samples result in a somewhat variable assay.
G-LISA assays are recommended for simple, reproducible quantification of results.
Provides a simple numeric readout
Reagents required that are not in the kits
The G-LISA assay provides all reagents necessary to perform 96 activation assays. Wells can be detached and used individually.