Millipore Technical Publications |
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Guava Tunel AssayFor quantifying mid- to late stage apoptosis |
| Assess late-stage apoptosis One of the hallmarks of late-stage apoptosis, or programmed cell death, is the fragmentation of nuclear chromatin. This generates DNA strands with exposed 3’-hydroxyl ends, which are enzymatically labeled in the classic TUNEL assay. Although regular TUNEL analysis is complicated and difficult, Guava’s flow cytometry kit makes this classic assay easy and accessible to everyone. Quantitative fluorescence assay The Guava TUNEL Assay detects apoptosis-induced DNA fragmentation through a quantitative fluorescence assay. Terminal deoxynucleotidyl transferase (TdT) catalyzes the incorporation of bromo-deoxyuridine (BrdU) residues into the fragmented nuclear DNA at the 3’-hydroxyl ends. A TRITC-conjugated anti-BrdU antibody then labels these DNA fragments. The assay distinguishes two populations: non-apoptotic cells (TUNEL-negative) and apoptotic cells (TUNEL-positive). Click the PDF above for the full document. | |
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