Molecular Probes™ CellEvent™ Caspase-3/7 Green Flow Cytometry Assay Kit

Catalog No.	Quantity
C10740	20 Assays
50-113-8070	100 Assays

Catalog No. C10740 Supplier Molecular Probes™ Supplier No. C10740

Description

The CellEvent™ Caspase-3/7 Green Flow Cytometry Assay Kit enables the flow cytometric detection of activated caspase-3 and caspase-7 in apoptotic cells.

The kit includes the novel fluorogenic substrate CellEvent™ Caspase-3/7 Green Detection Reagent, as well as SYTOX™ AADvanced™ Dead Cell Stain.

• Caspase-3/7 specific—includes the recognition sequence for activated caspase-3 and caspase-7
• Easy identification—clearly identify live, dead, and apoptotic cell populations
• Quick analysis—washing and fixation is not required
• Multicolor compatibility—combine with other dyes excitable by the 488 nm laser or other lasers

CellEvent™ Caspase-3/7 Green Detection Reagent is a cell-permeant reagent that 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, fluorogenic signal that has absorption/emission maxima of ∼511/533 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: 20 Assays
• Format: Kit
• Product Type: Caspase Assay Kit
• Content And Storage: Contains 1 vial CellEvent™ Caspase 3/7 Green Detection Reagent, 1 vial SYTOX AADvanced dead cell stain, and 1 vial of DMSO.
  
  Store all vials in the kit upright, at or below -20deg;C, desiccated, and protected from light.
• Flow Cytometer Laser Lines: 488
• Excitation/Emission: SYTOX AADvanced: 546/647
  CellEvent Caspase 3/7 Green Detection Reagent: 511/533
• Shipping Condition: Wet Ice
• Product Line: CellEvent
• Detection Method: Fluorescence
• For Use With (Application): Flow Cytometry
• For Use With (Equipment): Attune™ Acoustic Focusing Cytometer, Flow Cytometer
• Label or Dye: CellEvent™ Caspase 3/7 Green Detection Reagent, SYTOX AADvanced dead cell stain
• Label Type: Other Label(s) or Dye(s)

Frequently Asked Questions (FAQs)

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.

What are the advantages of flow cytometry?

-Measures data from single cells.
-Data are obtained for a large number of cells, generating a rich statistical analysis of cell populations.
-Because single cells are measured, it will reveal heterogeneity within a population.
-With the ability to multiplex, small sub-populations can be identified.
-Thousands of cells can be analyzed rapidly.
-It is ideally suited for blood samples and other cells in suspension.
-Data can be re-analyzed multiple times after acquisition.
-Flow cytometry files (FCS) can be archived.

What kinds of applications can I run on a flow cytometer?

There are several applications, some of which include immunophenotyping, cell cycle analysis, apoptosis assays such as annexin V staining, CellEvent Caspase-3/7 assay, and TUNEL assay, cell viability, proliferation assays such as CellTrace assay and Click-iT EdU assay, measurements of mitochondrial potential with MitoProbe assays, and cell counting using counting beads.

For Research Use Only. Not for use in diagnostic procedures.