04/11/11 - Multi-Parameter Apoptosis Assay Kit

Phenotypic characterization of different cell death parameters at a single-cell level

Multi-Parameter Apoptosis Assay Kit employs FITC-conjugated Annexin V as a probe for phosphatidylserine on the outer membrane of apoptotic cells, TMRE as a probe for mitochondrial membrane potential, 7-AAD as an indicator of membrane permeability/cell viability, and Hoechst Dye to demonstrate nuclear morphology.

The kit allows phenotypic characterization of different cell death parameters at a single-cell level.

The assay can be adapted to high content screening with appropriate equipment. The reagents provided in the kit are sufficient to run 100 samples when using flow cytometry, or 500 samples when using a 96-well plate format.

Multi-parameter with KA1335

 Immunofluorescence

Staurosporine induces apoptosis in Jurkat cells, as measured by

• nuclear morphology,
• a decrease in mitochondrial membrane potential, and
• an increase of Annexin V FITC positive cells.

Panels A-C show same field of cells from the control group / Panels D-F show the same field of cells from the staurosporine-treated group

Hoechst staining shows that most control cells have round and intact nuclei (Panel A) whereas staurosporine-treated cells mostly have condensed and fragmented nuclei (Panel D).

TMRE staining reveals that most control cells have undisrupted mitochondrial membrane potential (Panel B) whereas staurosporine-treated cells mostly have diminished membrane potential and were not stained (Panel E).

Most control cells are Annexin V negative (Panel C) whereas cells treated with staurosporine are mostly Annexin V positive (Panel F), indicating cells are undergoing apoptosis.