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Detection of Apoptosis by CaspSCREEN™ Flow Cytometric Analysis Kit
CaspScreen™, non-adherent cells only
Description:
CaspScreen™, non-adherent cells only
Trade Name:
- CaspSCREEN
- Chemicon (Millipore)
Qty/Pk:
25 assays
Product Overview:
Activation of ICE-family proteases/caspases initiates apoptosis in mammalian cells. The CaspSCREEN™ Flow Cytometric Apoptosis Detection Kit provides a simple and convenient means for detecting activation of caspases by flow cytometry in intact cells. The assay utilizes a substrate that contains two aspartate residues linked to rhodamine 110 (D2R), a reported substrate for members of capase family proteases. The rhodamine 110 conjugated substrate is non-fluorescent, however, upon cleavage of the substrate by cellular caspases or apoptosis-related proteases, the released rhodamine 110 gives rise to fluorescence that can be measured at excitation of 488 nm and emission of 530 nm. As the D2R substrate is more cell-permeable than other fluorometric caspase substrates, apoptosis can easily be measured in intact cells by flow cytometry.
Key Applications:
- Flow Cytometry
- Activity Assay
Species Reactivity:
Mammals
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Components:
- D(2)R - 25 μL
- DTT (1 M): 75 μL
- Incubation Buffer: 7.5 mL
Brand Family:
Chemicon
Storage Conditions:
We recommend storing reagents at -20°C upon arrival. (Store Incubation Buffer at 4°C after opening). The performance of this product is guaranteed for 6 months.
Detection Methods:
Fluorescent
Protein/Isoform Description:
Cysteine aspartyl proteases related to the C. elegans CED-3 death protein comprise the caspase family. All are expressed as proenzymes which are activated by proteolysis. With respect to their roles in apoptosis, Caspases can be subdivided into initiator (Caspases 8, 9, 10) and effector (Caspases 3, 6, 7) caspases, depending on whether they are activated by receptor clustering (initiator) or by mitochondrial permeability transition (effector).
Effector caspases, most notably Caspase 3, cleave numerous substrates to effect the morphological changes associated with apoptosis. Among Caspase 3 substrates are DFF45/ICAD, which frees up the DNAse subunit of DFF to cause chromatin degradation, as well as gelsolin, PAK2, D4GDI, all of which are involved in cytoskeletal organization, nuclear lamins and PARP. The significance of PARP cleavage is not clear, but it is an excellent marker for caspase activation and the presumption of ongoing apoptosis.
Effector caspases, most notably Caspase 3, cleave numerous substrates to effect the morphological changes associated with apoptosis. Among Caspase 3 substrates are DFF45/ICAD, which frees up the DNAse subunit of DFF to cause chromatin degradation, as well as gelsolin, PAK2, D4GDI, all of which are involved in cytoskeletal organization, nuclear lamins and PARP. The significance of PARP cleavage is not clear, but it is an excellent marker for caspase activation and the presumption of ongoing apoptosis.
Product Name:
CaspScreen™, non-adherent cells only
Materials Required but Not Delivered:
· Microcentrifuge and 1.5 mL Microcentrifuge tubes
· 37°C water bath or incubator
· Flow Cytometer
· 37°C water bath or incubator
· Flow Cytometer