Multiplex Apoptosis and Necrosis

Multiplex Apoptosis and Necrosis

Apoptosis is an active, programmed process of autonomous cellular dismantling that avoids eliciting inflammation. In apoptosis, phosphatidylserine (PS) is transferred to the outer leaflet of the plasma membrane. As a universal indicator of the initial/intermediate stages of cell apoptosis, the appearance of phosphatidylserine on the cell surface can be detected before morphological changes are observed.

Necrosis is characterized as a passive, accidental cell death resulting from environmental perturbations with uncontrolled release of inflammatory cellular contents. Loss of plasma membrane integrity represents a straightforward approach to demonstrate late stage apoptosis and necrosis.

Cell Meter™ Apoptotic and Necrotic Detection Kits are a set of tools for monitoring cell viability. The kits are optimized to simultaneously detect cell apoptosis, necrosis and healthy cells with a flow cytometer or fluorescence microscope. The phosphatidylserine (PS) sensor used in Kit 22843 has deep red fluorescence (Ex/Em = 630/660 nm) upon binding to membrane PS. Membrane-impermeable Nuclear Green™ DCS1 (Ex/Em = 490/525 nm) is used to label the nucleus while CytoCalcein™ Violet 450 (Ex/Em = 405/450 nm) is provided for labeling live cell cytoplasm.

Key Features of Cell Meter™ Apoptotic and Necrotic Detection Kits (pg 15-16 of this catalog):

• Multiplexing capability, triple colors for the simultaneous detection of multiple cellular events.
• Robust, a mix and read format.
• Convenient, compatible with common filter sets.

The fluorescence images of Jurkat cells showing cells that are alive (blue, stained by CytoCalcein™ Violet 450, Cat# 22012), apoptotic (green, stained by Apopxin™ Green), and necrotic (red, indicated by 7-AAD staining, Cat# 17501). The fluorescence images of the cells were taken with Olympus fluorescence microscope in the Violet, FITC and TRITC channel respectively. Individual images taken in each channel from the same cell population were merged as shown above. B. Non-induced control cells. C: Triple staining of staurosporine-induced cells (1 μM staurosporine treatment for 3 hours.)

The detection of apoptosis and necrosis with Kit 22843. Binding activity of Apopxin™ Deep Red to phosphatidylserine in Jurkat cells. The fluorescence imaging demenstrated that live cells (blue) were stained by CytoCalcein™ Violet 450 (Cat# 22012), apoptotic cells (red) were stained by Apopxin™ Deep Red, and necrotic cells (green) were stained by Nuclear Green™ DCS1 (Cat# 17550). Apoptosis was induced by 1 μM staurosporine for 3 hours. The fluorescence images of the cells were taken with Olympus fluorescence microscope using the Violet, Cy5® and FITC channel respectively. Individual images taken in each channel from the same cell population were merged as shown above. A: Non-induced control cells; B: Triple staining of staurosporine-induced cells.

Product Ordering Information:

Cat# 22840 Cell Meter™ Apoptotic and Necrotic Detection Kit *Triple Fluorescence Colors*
Cat# 22843 Cell Meter™ Apoptotic and Necrotic Detection Kit *Triple Fluorescence Colors*