Apoptosis is essential in many physiological processes. Several methods have been described to identify apoptotic cells. Endonucleolysis is considered as the key biochemical event of apoptosis, resulting in double-stranded, low molecular weight DNA fragments as well as single strand breaks in high molecular weight DNA. Those DNA strand breaks can be identified by labeling free 3’-OH termini with modified nucleotides in an enzymatic reaction. Terminal deoxynucleotidyl transferase (TdT) can catalyze DIG-dUTP to free 3’-OH DNA ends. The DIG-dUTP is first react with biotin-conjugated anti-digoxin antibody, which subsequently associate with streptavidin-AP(SABC-AP). Combined with BCIP/NBT Chromogenic Reagent, apoptotic cells stain Purple-blue analyzed under microscopy.
Component A: Labeling Buffer; Component B: TdT (20X); Component C: DIG-dUTP (20X); Component D: Blocking Reagent; Component E: Biotin-conjugated anti-digoxin antibody; Component F: SABC-AP (100X); Component G: Protein K (200X); Component H: BCIP/NBT Chromogenic Reagent (20X); Component I: Antibody Dilution; Positive Control
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