Sulforhodamine Caspase Detection Kit
Other caspase detection assays on the market work through substrate cleavage mechanisms or immunohistochemical principles. APO LOGIX reagents work by active caspase inhibition. Since apoptosis is arrested upon the binding of the reagent to the active caspase heterodimer, the apoptotic cell population does not diminish over time.
Easy to Use:
Add reagent directly to cells. No special buffer or media
needed. No preparation of cell lysates required. Simple
wash procedure.
Quick:
Incubate for one hour, wash twice, and measure. No time
course studies necessary.
Adaptable:
Works in diverse cell lines: human, rodent, Drosophila.
Can be performed in conjunction with Annexin staining,
TUNEL, antibody staining, or with other APO LOGIX
reagents on the same population of cells. Permits high
through-put screening. Protocol can be adapted for ex
vivo as well as in situ experiments.
Non-Cytotoxic:
Arrests further apoptotic activity via caspase inhibition.
Cell Permeable:
Permits direct visualization of cytosolic apoptotic events.
Compatible with Multiple
Fluorometric Modalities:
Fluorescence microscopy. 96-well fluorescence plate reader.
Reliable:
Yields both quantitative and qualitative results. Gives
strong signal with little background noise.
Optimizable:
Cell Technology offers free technical support to help
theresearcher optimize the application of the APO LOGIX
reagents to particular experimental contexts.
Comprehensive:
APO LOGIX reagents mark activity across the range of
caspase proteins. Our poly-caspase assay is useful for visualizing
general caspase activity.
APO LOGIX
Sulforhodamine caspase detection kits label active caspases
in living cells that are undergoing apoptosis. Cell
Technology's probes utilize Sulforhodamine (SR) labeled
peptide fluoromethyl ketone (FMK) caspase inhibitors
(SR-peptide- FMK). These SR-peptide-FMK are both cell
permeable and non-cytotoxic during the course of the
assay, thus allowing the detection of active caspases in living
cell systems.
Assay Principle:
When added to a population of cells undergoing apoptosis,
the SR-peptide-FMK probe enters each cell and covalently
binds to the reactive cysteine residue on the large subunit
of the caspase heterodimer, thereby the probe is
sequestered and accumulates inside the cell. At the same
time it inhibits further caspase activity. The remaining
unbound reagent is easily washed away. Cells with the
bound SR-peptide-FMK can be detected by flow cytometry,
fluorescence microscope or a 96 well fluorescence plate
reader. The SR-peptide-FMK reagent excites at 550 nm
and has a maximum emission range of 590-600 nm.
Cell Technology, Inc., 2010 E. Hennepin Ave - Suite 203 , Minneapolis, MN 55413. Tel: 612-331-4704; Fax: 612-378-2319 .