The Hoechst 33342/Propidium Apoptosis Assay Kit provides a rapid and convenient assay for apoptosis based upon fluorescence detection of the compacted state of the chromatin in apoptotic cells. The kit contains ready-to-use solutions of the blue fluorescent Hoechst 33342 dye (excitation/emission at 350/461 nm, when bound to DNA), which stains the condensed chromatin of apoptotic cells more brightly than the chromatin of normal cells, and the red-fluorescent propidium iodide dye (excitation/emission at 535/617 nm, when bound to DNA), which is permeant only to dead cells. The staining pattern resulting from the simultaneous use of these dyes makes it possible to distinguish normal, apoptotic and dead cell populations by flow cytometry. The 351 nm line of an argon-ion laser or other suitable UV source is required for excitation of the Hoechst 33342 dye, whereas propidium iodide may be excited with the 488 nm line of an argon-ion laser. We have optimized this assay using Jurkat cells, a human T-cell leukemia clone, treated with camptothecin to induce apoptosis. Some modifications may be required for use with other cell types.
Flow Cytometry, Fluorescence Microscopy
50tests ; 100tests ; 200 tests
Upon receipt, store the kit at 4°C, protected from light. The components of the kit should be stable in storage for at least 6 months. Hoechst 33342 and propidium iodide are light sensitive. They may be handled in normal room light, but avoid prolonged exposure to light.
Celltechgen provides high-quality reagents and materials for research use only. Propidium iodide and Hoechst 33342 are known or suspected mutagens and should be used with appropriate precautions. For proper handling of potentially hazardous chemicals, please request the Safety Data Sheet (SDS) provided for the product.