Promega's homogeneous caspase assays are highly reproducible as measured by Z' factors.
Z' factor values are statistical indicators of the dynamic range and variability of assay results. Assays that produce Z' factor values of 0.5 or greater are considered well suited for high throughput screening.
|
Z' Values |
# Wells |
|
| Caspase-Glo® 3/7 Assay |
0.92 |
96 |
The Caspase-Glo 3/7 Assay can produce excellent Z′-factor values.
Z′-factor value = 0.92 for this assay.
|
| Caspase-Glo® 8 Assay |
0.957 |
48 |
The Caspase-Glo 8 Assay can produce excellent Z′-factor values.
Z′-factor Value = 0.957 for this assay.
|
| Caspase-Glo® 9 Assay |
0.903 |
48 |
The Caspase-Glo 9 Assay can produce excellent Z′-factor values.
Z′-factor Value = 0.903 for this assay.
|
| Caspase-Glo® 2 Assay |
0.89 |
384 |
Z′-factor analysis for Caspase-Glo 2 Assay.
Z′-factor values (15) for the Caspase-Glo® 2 Assay were calculated using recombinant caspase-2 (5U, 0.5U, and 0.05U/ml) and a no-caspase blank. Caspase-2 was diluted in 10mM HEPES (pH 7.0), 2mM DTT, 0.1% Prionex® as a carrier. Assays were performed in a total volume of 20µl in a 384-well plate. Luminescence was recorded on a BMG PHERAstar luminometer at 30 minutes. Z′-factor values ranged from 0.85 to 0.91 for this assay. One unit of caspase-2 is defined as the amount of enzyme required to cleave 1pmol of substrate/minute at 30°C using 200 µM Ac-LEHD-pNA as substrate. In this case, 1 unit of caspase-2 is equivalent to 10ng protein.
|
| Caspase-Glo® 6 Assay |
0.86 |
384 |
Z′-factor value analysis of the Caspase-Glo® 6 Assay.
Z′-factor values were calculated using 1U/ml and 0.5U/ml recombinant caspase-6 (BIOMOL International) and a no-caspase blank. Assays were performed in a 384-well plate using the Caspase-Glo® 6 Assay. Luminescence was recorded as relative light units (RLU) on a BMG FLUOstar multimode reader 30 minutes after adding the Caspase-Glo® 6 Reagent.
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