What is caspase-3 activity assay?
The Caspase-3 Activity Assay Kit is a fluorescent assay that detects the activity of caspase-3 in cell lysates. It contains a fluorogenic substrate (N-Acetyl-Asp-Glu-Val-Asp-7-amino-4-methylcoumarin or Ac-DEVD-AMC) for caspase-3.
What is caspase activity assay?
The caspase activity assay is designed for use with multiwell plate formats using either purified enzyme or cells in culture and can be multiplexed with other cell-based assays. The assay delivers excellent Z´-factor values in cell and purified enzyme models.
How is caspase-3 activity measured?
The Caspase-3 assay protocol is based on the formation of the chromophore p-nitroaniline (p-NA) by cleavage from the labeled substrate DEVD-pNA. The p-NA can be quantified using a spectrophotometer or a microtiter plate reader reading absorbance at 400 or 405 nm.
What is a limitation of the caspase-3 assay?
Additionally, the main limitations for our multiplexing protocol are that caspase-3/7 activity can determined but not quantified, and the resulting lysate cannot be used for downstream assays (i.e. Western blots or gene expression).
How is caspase activity measured?
Caspase activity was determined by quantifying fluorescence images after excitation at 380 nm and emission centered at both 545 nm (yellow fluorescence, i.e. FRET) and 450 nm (blue fluorescence, i.e. conventional excitation) were captured.
What is caspase 3 7 assay protocol?
Caspase-Glo® 3/7 Assay Protocol. The Caspase-Glo® 3/7 Assay is a luminescent assay that measures caspase-3 and -7 activities in purified enzyme preparations or cultures of adherent or suspension cells. The assay provides a proluminescent caspase-3/7 substrate, which contains the tetrapeptide sequence DEVD.
How many caspase activity assays can I perform with caspase-Glo® reagent?
G8090 provides sufficient reagents to perform 25 caspase activity assays; Cat.# G8091, 100 assays; Cat.# G8092 and G8093, 1,000 assays. Using 25μl of Caspase-Glo® Reagent per assay in a 384-well format, Cat.#
What is the role of caspase-3 in apoptosis?
Caspase-3 activity assay Caspases play important roles in inducing apoptosis in which caspase-3 is an executive molecule. Caspase-3 has many functions in various pathways via which apoptosis signals transduce. Caspase-3 normally exists in cytoplasm in form of pro-enzyme. In initiation stage of apoptosis, caspase-3 is activated.
What is Apo-one homogeneous caspase-3/7?
The Apo-ONE® Homogeneous Caspase-3/7 Assay provides the necessary reagents for fast and sensitive measurement of active caspase-3 and -7 in a homogeneous format. The assay includes a profluorescent caspase-3/7 consensus substrate, rhodamine 110 bis-(N-CBZ- l -aspartyl- l -glutamyl- l…