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Comprehensive Analysis of Necrotic, Apoptotic, and Live Cells Using Multicolor Staining

This protocol allows simultaneous measurement of necrotic, apoptotic dead, and live cells. Treat cells with DNAse to nick cells, then incorporate FITC-UTP with TdT for apoptotic evaluation. Fix cells and permeabilize, followed by incorporating B-UTP with TdT and PE-Avidin for necrotic cell assessment. Simultaneously evaluate dead, necrotic, and apoptotic cells by flow cytometry.

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Comprehensive Analysis of Necrotic, Apoptotic, and Live Cells Using Multicolor Staining

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  1. Simultaneous Measurement of Necrotic, Apoptotic Dead and Live Cells Necrotic/Apoptotic Dead Treat Cells with DNAse to nick cells Incorporate FITC-UTP with TdT Apoptotic Live Fix cells and permeabilize Incorporate B-UTP with TdT then PE-Avidin

  2. SIMULTANEOUS EVALUATION OF DEAD, NECROTIC AND APOPTOTIC CELLS 0 1 2 3 4 10 10 10 10 10 0 256 512 768 1024 FITC UTP -> FSC -> TDT48015 TDT48015

  3. LIGHT SCATTER ANALYSIS OF DEAD CELLS AND VIABLE APOPTOTIC CELLS R4 R3 gated R1 A B PE-dUTP SSC R1 R2 Fl-12-dUTP FSC gated R2 gated R3 C D SSC SSC FSC FSC

  4. FOUR COLOR DETECTION OF SURFACE ANTIGENS, APOPTOSIS AND DEAD CELLS DEAD CELLS DEAD CELLS PE-CD19 APOPTOSIS TC-CD33 APOPTOSIS

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