Experience Improved Selectivity for Caspase-8 & Caspase-9
Caspase-Glo® 8 Assay uses a luminogenic substrate containing the LETD sequence, and Caspase-Glo® 9 Assay uses LEHD sequence, both of which have been shown to be selective for their respective caspase enzymes (1,2). The assays include an optional proteasome inhibitor (MG-132), which when added to the Caspase-Glo Reagent, significantly reduces nonspecific background in cell-based assays.
Comparison of the Caspase-Glo 8 Assay with and without the proteasome inhibitor, MG-132. Note the improved signal:noise ratio when MG-132 is included as well as the improved Z'-factor, an indication of assay performance widely used in screening applications. Data was generated in 384-well plates.
Caspase-Glo 8 & Caspase-Glo 9 Assays are homogeneous luminescent assays that selectively measure caspase-8 or caspase-9 activity. The assays provide proluminogenic substrates in a buffer system optimized for caspase activity, luciferase activity and cell lysis. The addition of a single Caspase-Glo Reagent in an "add-mix-read" format results in cell lysis, followed by caspase cleavage of the substrate and generation of a "glow-type" luminescent signal. The signal generated is proportional to the amount of caspase activity present. The Caspase-Glo Reagent relies on the properties of the proprietary thermostable luciferase, Ultra-Glo™ Recombinant Luciferase, which generates a stable luminescent signal and improves performance across a wide range of assay conditions.
For additional information on these assays, please request or download the Technical Bulletins from the Promega web site: Caspase-Glo® 8 Assay or Caspase-Glo® 9 Assay
SIDEBAR: If you are interested in trialing a new Caspase-2 or Caspase-6 assay currently in development, please contact Promega for information.
Neal Cosby, PhD, Strategic Marketing Manager neal.cosby@promega.com
References
1. Thornberry, N.A., Chapman, K.T. and Nicholson, D.W. (2000) Methods Enzymol. 322, 100–10.
2. Garcio-Calvo, M. et al. (1999) Cell Death Differ. 6, 362–9.
To learn more about the Caspase-Glo Assays from Promega: www.promega.com/drugdiscovery
 
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PROTEASOME ASSAYS MADE EASY
 Promega recently launched a series of assays for the detection of chymotrypsin-like protease activity of the proteasome complex in cultured cells (Proteasome-Glo™ Cell-Based Assay) and for the measurement of the three proteolytic activities associated with the proteasome in an enzyme-based format (Proteasome-Glo™ 3-Substrate System).
These homogeneous bioluminescent assays feature simple "add-mix-measure" protocols and greater sensitivity with linearity over 3 logs of cell number or 4 logs of proteasome concentration, and are ideally suited for measuring the chymotrypsin-like activity of the proteasome in cultured cells or screening of inhibitor libraries and monitoring proteasome-regulated protein degradation in cells.
To learn more about the Proteasome-Glo Assays from Promega: www.promega.com/drugdiscovery
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