Vybrant® FAM Caspase-3 and -7 Assay Kit, for flow cytometry

Catalog number: V35118

Molecular Probes™  Related applications: Cell Viability, Proliferation & Function | Cellular Imaging | Flow Cytometry | Target-Based ADME⁄Tox Assays

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Description

The Vybrant™ FAM Caspase-3 and -7 Assay Kit employs a novel approach to detect active caspases that is based on a fluorescent inhibitor of caspases (FLICA™) methodology, essentially an affinity label. The reagent associates a fluoromethyl ketone (FMK) moiety, which can react covalently with a cysteine, with a caspase-specific amine acid sequence. For caspase-3 and -7, this recognition sequence is aspartic acid-glutamic acid-valine-aspartic acid (DEVD). A fluorescein group is attached as a reporter. The FLICA reagent is thought to interact with the enzymatic reactive center of an activated caspase via the recognition sequence, and then to attach covalently through the FMK moiety. The FLICA inhibitor is cell permeant and noncytotoxic. Unbound FLICA molecules diffuse out of the cell and are washed away; the remaining green-fluorescent signal is a direct measure of the amount of active caspase that was present at the time the inhibitor was added. This kit includes the FLICA reagent specific for caspase-3 and -7, Hoechst 33342 stain, and propidium iodide stain, which allows the simultaneous evaluation of caspase activation, membrane permeability, and cell cycle by flow cytometry.

View a selection guide for all apoptosis assays for flow cytometry.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Format: Tube(s)
Label or Dye: Propidium iodide, FAM™, Hoechst 33342
Product Size: 25 assays
Detection Method: Fluorescent
Number of Reactions: 25
For Use With (Equipment): Flow Cytometer
Excitation⁄Emission (nm): PI: 535⁄617, Hoechst 33342: 350⁄461, FAM-DEVD-FMD: 388⁄530
Shipping Condition: Room Temperature

Contents & storage

Contains 1 vial FAM-DEVD-FMK caspase-3 and -7 reagent (FLICA reagent, lyophilized solid), 1 vial of Hoechst 33342 (400 µL, 1 mM in water), 1 vial of PI (1 mL, 250 µg/mL in water), 1 vial of DMSO (0.5 mL), 1 bottle of apoptosis fixative solution (10% formaldehyde with methanol in PBS, 6 mL), and 1 bottle of 10x apoptosis wash buffer.

Store in refrigerator (2–8°C) and protect from light.

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