PF032 Sigma-AldrichAnnexin V-FITC Apoptosis Detection Kit

Figure 1: Principle of the Assay

Figure 2: Flow Cytometry

A sample cytogram is shown above. Viable cells do not bind Annexin V-FITC or propidium iodide as reflected in the lower left-hand quadrant of the dot plot. Early apoptotic cells with exposed PS but intact cell membranes bind Annexin V-FITC but exclude propidium iodide. Fluorescence from this population is reported in the lower right-hand quadrant. Necrotic or apoptotic cells in terminal stages will be both Annexin V-FITC and propidium iodide positive and will be reported in the upper right-hand quadrant. A small percentage of normal cell death should be expected in routine cultures of untreated cells.

Figure 3: RAPID Annexin V-FITC binding is identical to results with Conventional Protocol

Jurkat cells were incubated with 0.5 µg/ml actinomycin D for either 4.5 (Middle) or 19 (Bottom) h. Untreated shown in the TOP panel of each section. Annexin V-FITC was then bound in either binding buffer (using the Conventional protocol) or directly in either RPMI, or DMEM or McCoy's 5A (RAPID Protocol). The log of both PI and FITC fluorescence is reported.

Figure 4: RAPID Annexin V-FITC binding is identical to results with Conventional Protocol

Apoptosis was induced by 30 min of UV irradiation followed by either 4 (Middle) or 18 (Bottom) h incubation at 37°C. Untreated cells are shown in the Top panel of each section. Annexin V-FITC was then bound in either binding buffer (using the Conventional protocol) or directly in either RPMI, or DMEM or McCoy's 5A (RAPID Protocol).