Flow Cytometry apoptosis detection
two reagents for apoptosis analysis...
Biostatus offer two flow cytometry approaches to the study of apoptosis:-
1. See apoptotic populations using sub-G1 cell cycle analysis with DRAQ5™
2. Positively discriminate LIVE and apoptotic cells using APOPTRAK™
1. Sub-G1 cell cycle analysis as a marker of apoptotic DNA fragmentation
With DRAQ5™ you can generate cell cycle data and measure the presence of a sub-G1 fraction. This indicates DNA fragmentation resulting from apoptosis.
Snyder et al (2004) used this strategy when studying the potential for a peptide treatment of advanced peritoneal carcinomatosis. Cultured Namalwa human B-lymphoma cells were directly stained with DRAQ5™ and analysed by flow cytometry.
Due to the far-red emission of DRAQ5 this could also be preceded by a immunochemical staining for antigens to determine phenotype for apoptotic cells in primary / heterogeneous samples. See cell cycle analysis for more information on this topic. See the DRAQ5 protocol.
2. Tracking apoptosis - discriminating intact, leaky & early apoptotic cells
See the example protocol for use of APOPTRAK™ in flow cytometry. Navigate to the main product section on APOPTRAK™ for further information.
Uptake of APOPTRAK in B cell lymphoma carrying a low burden of apoptotic cells. Cells were co-stained with FITC-Annexin V:
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