| Western blot |
Luminescence |
Low |
Technical simplicity, standard instrumentation, gold standard for many studies |
Labor intensive, highly variable, difficult to standardize |
| Cellular image-based assays |
| LICOR |
Fluorescence |
Medium |
Standard instrumentation |
Low resolution imaging, typically singleplex, as labor intensive as HCI |
| High Content Imaging (HCI) |
Fluorescence |
Medium to high |
High resolution imaging, population analysis, easy to multiplex |
Labor intensive, requires specialized instrumentation, special software for analysis |
| Single-parameter well-based assays |
| Enzyme-linked immunosorbent assay (ELISA) |
Colorimetric, luminescence, fluorescence |
Low to medium |
Good dynamic range, quantitative, robust, cost-efficient |
Labor intensive: multistep preparation process, low-throughput |
| Electrochemiluminescence (ECL) |
Luminescence |
Medium to high |
Large dynamic range and sensitivity, potential for multiplexing, robust |
Labor intensive, costly, requires specialized instrumentation |
| Time-resolved Fluorecence Energy Resonance Transfer (TR-FRET) |
Fluorescence |
High |
Homogeneous, high-throughput, good dynamic range, robust, cost-efficient |
Sensitive to some conditions and reagents, limitations imposed by distance for FRET |
| Amplified Luminescent Proximity Homogenous Assay Screen (AlphaScreen®) |
Fluorescence |
High |
Homogeneous, high-throughput, large dynamic range and sensitivity, robust in variable conditions |
Requires specialized instrumentation, more expensive then FRET |
