Invitrogen™ CellEvent™ Caspase-3/7 Detection Reagents

Description

• CellEvent Caspase-3/7 Green and CellEvent Caspase-3/7 Red detection reagents are novel fluorogenic substrates for activated caspase-3/7 in live cells that can be fixed and multiplexed with antibody detection workflows such as immunofluorescence (IF) and immunocytochemistry (ICC)
• Activation of caspase-3 is an essential event during apoptosis, making these optimized reagents for analysis of apoptotic cells
• CellEvent Caspase-3/7 Red Detection Reagent can also be used to detect apoptosis in GFP expressing cells
• Features of these indicators include: • Optimized caspase-3/7 substrate for apoptosis analysis • Easy to use—add and read; no cell lysis required • Use for time course measurements; easily select the time point of interest • Compatible with both live cell fluorescence-imaging and formaldehyde-based fixation methods • Multiplex enabled—combine with other fluorescent reagents to confirm apoptosis in the same cell or cell population CellEvent™ Caspase-3/7 Green Detection Reagent : Absorption/emission maxima of ∼502/530 nm
• Detectable with FITC/GFP traditional filter set
• CellEvent™ Caspase-3/7 Red Detection Reagent : Absorption/emission maxima of ∼590/610 nm
• Detectable with traditional Texas red filter set
• CellEvent™ Caspase-3/7 Red Detection Reagent and CellEvent™ Caspase-3/7 Green is provided as Dry down powder
• Just add 100μL of PBS to dry down powder to generate 100X Stock solution
• Dilute this 100X stock solution 1:100 in complete media and stain cells for 30-60 minutes and analyze with fluorescence microscope
• CellEvent™ Caspase-3/7 Red Detection Reagent is also provided as already prepared DMSO solution
• DMSO solution is 400X stock solution, which is ready to use
• Just dilute in growth media 1:400 and stain the cells for 30 to 60 minutes
• To prevent the apoptotic cells from washing away, no wash is needed at the end of staning
• Stained cells can be analyzed live, or can be fixed with 4% Paraformaldehyde solution such as Image-IT™ Paraformaldehyde 4% in 0.1M Phosphate Buffer pH 7.4, Cat
• No
• I28800
• These fixed cells can be further processed for ICC workflow
• Substrate specifics CellEvent Caspase-3/7 Detection Reagents are a four-amino acid peptide (DEVD) conjugated to a nucleic acid-binding dye
• The DEVD peptide sequence is a cleavage site for caspase-3/7, and the conjugated dye is non-fluorescent until cleaved from the peptide and bound to DNA
• Principles of action CellEvent Caspase-3/7 Green Detection Reagent is intrinsically non-fluorescent as the DEVD peptide inhibits the ability of the dye to bind to DNA
• However, after activation of caspase-3/7 in apoptotic cells, the DEVD peptide is cleaved, enabling the dye to bind to DNA and produce a bright, fluorogenic response
• Simple three-step protocol To use CellEvent Caspase 3/7 Detection Reagent, simply add it to cells, incubate 30–60 minutes, and visualize
• Apoptotic cells with activated caspase-3/7 will have bright green or red nuclei, while cells without activated caspase 3/7 will have minimal fluorescent signal
• Assay versatility allows for live cell detection and fixation This robust assay enables the examination of caspase-3/7 activation in live cells
• Furthermore, since wash steps are not necessary for detection, fragile apoptotic cells typically lost during wash steps are preserved
• Importantly, the fluorescent signal from CellEvent Caspase 3/7 Detection Reagent can survive fixation and permeabilization
• This allows for the flexibility to perform end-point assays and probe for other proteins of interest using immunocytochemistry.