Invitrogen™ CellEvent™ Caspase 3/7 Green Ready Flow™ Reagent

Catalog No.	Quantity
R37167	120 assays

Catalog No. R37167 Supplier Invitrogen™ Supplier No. R37167

Description

CellEvent™ Caspase-3/7 Green Ready Flow™ Reagent enables the flow cytometric detection of activated caspase-3 and caspase-7 in apoptotic cells.

The product includes 2 reagents: the novel fluorogenic substrate CellEvent™ Caspase-3/7 Green Detection Reagent and SYTOX™ AADvanced™ Dead Cell Stain.

• Stable at room temperature
• Convenient, ready-to-use format—no need to dilute, weigh, or pipette
• Exceptionally bright green fluorescence upon binding to nucleic acids
• Rapid staining of apoptotic cells without wash steps

CellEvent Caspase-3/7 Green Ready Flow Reagent is a cell-permeant reagent that detects caspase 3/7 activity using flow cytometry. Add 2 drops from the dropper bottle directly into 1 x 10⁶ cells, incubate, and analyze.

CellEvent Caspase-3/7 Green Ready Flow Reagent consists of a four-amino acid peptide (DEVD) conjugated to a nucleic acid-binding dye. During apoptosis, caspase-3 and caspase-7 proteins are activated and able to cleave the caspase 3/7 recognition sequence encoded in the DEVD peptide. Cleavage of the recognition sequence and binding of DNA by the reagent labels the apoptotic cells with a bright, fluorescent signal having absorption/emission maxima of ∼502/530 nm. When used together with the SYTOX AADvanced Dead Cell Stain, apoptotic cells can be easily discriminated from live and necrotic cells.

Because no single parameter defines apoptosis in all systems, we strongly suggest using a combination of different measurements for reliable detection of apoptosis.

Specifications

• Quantity: 120 assays
• Format: Dropper Bottle
• Product Type: Caspase 3/7 Reagent
• Content And Storage: 4 dropper bottles, store at room temperature
• Excitation/Emission: 504/523
• Color: Green
• Shipping Condition: Room Temperature
• Sub Cellular Localization: Nucleus
• Product Line: CellEvent™ Ready Flow
• Label or Dye: CellEvent™ Caspase 3/7 Green, SYTOX AADvanced Dead Cell Stain
• Label Type: Other Label(s) or Dye(s)

Frequently Asked Questions (FAQs)

I would like to label only 100 µL of sample of the same cell density using a Ready Flow product. The manual notes 2 drops per 1 mL (1 X 10E6 cells/mL). May I scale down for smaller volumes?

Yes, you may scale up or down as needed, but we recommend keeping the cell density the same. There are 41 µL per drop, so 2 drops is 82 µL (for the 1 mL of sample). You need to only scale down for this 100 µL volume, which would be 8.2 µL.

What is the concentration of dye/reagent in the Ready Flow products?

The amount of dye or reagent in the Ready Flow products is proprietary. If you need to consider the exact amount of dye or reagent for your experiment, each of the Ready Flow reagents is available as a standalone product.

Can I use the ReadyProbes reagents for flow cytometry?

This is not recommended. The ReadyProbes reagents were developed for imaging applications whereas the Ready Flow reagents were optimized for flow cytometry.

Can I use CellEvent Caspase-3/7 Green Detection Reagent with fixed samples?

No, it must be applied to live cell populations.

With the CellEvent Caspase-3/7 Green Detection Reagent, may I stain cells with nucleic acid stains?

Yes, you may use SYTOX AADvanced Dead Cell Stain that is included in the CellEvent Caspase-3/7 Green Ready Flow Reagent (Cat. No. R37167) and in the CellEvent Caspase-3/7 Green Flow Cytometry Assay Kit (Cat. No. C10427, C10740). You may also try using other nucleic acid stains in the red or far red channels for staining cells.

With the CellEvent Caspase-3/7 Green Detection Reagent, may I multiplex with fluorescent immunolabeling?

Yes. You can apply surface labels or other reagents. We recommend applying the surface labels or other reagents first.

Is the CellEvent Caspase-3/7 Green Detection Reagent fixable?

The CellEvent Caspase-3/7 Green Detection Reagent is not covalently attached to any cellular component and may be lost upon fixation. For flow cytometry applications, after applying the reagent, you should examine without washing or fixation. For imaging applications, the CellEvent product signal may be retained after fixation in 3.7% formaldehdye for 15 min.

Can I fix my CellEvent Caspase 3/7 - labeled cells?

For imaging applications, we recommend fixing with 3.7% formaldehyde at room temperature for 15 mins. After fixation, cells can be imaged for up to 24 hrs, but the signal will gradually degrade over time. For flow cytometry applications, fixation is not recommended.