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Apoptosis Assay Kit is based on standard setup that employs Annexin V-FITC conjugate and propidium iodide (PI). The flow cytometry test can discriminate intact cells (FITC-/PI-), apoptotic cells (FITC+/PI-) and necrotic cells (FITC+/PI+).
2. Binding Buffer
Quantity: 5 ml (10x concentrated).
Form: Concentrated filter-sterilized solution (0.1M HEPES/NaOH,
pH 7.4, 1.4M NaCl, 25mM CaCl2). Dilute 10X in deionized water
prior use.
JURKAT cells (human T cell leukemia) were treated with
200 ng/ml TRAIL/Apo2L (bacterially expressed extracellular
domain of human TRAIL, corresponding to amino acids 95-281)
for 6 hours or left untreated.
Detection of apoptotic cells was preformed by flow cytometry
using Apoptosis Assay Kit (Cat. No. KIT-ANXF).
Legend:
Green line - untreated and unstained cells
Red line - untreated and stained cells
Blue line - TRAIL-treated and stained cells.
A small proportion of untreated cells were identified as undergoing apoptosis (PS expressing, PI negative). In TRAIL-treated cells the number of early apoptotic cells (PS expressing, PI negative) as well as necrotic cells (PS expressing, PI positive cells) significantly increased.
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