TUNEL Apoptosis Detection Kit (DNA Fragmentation/Fluorescence Staining), BioAssay(TM)

Suitable for the detection of the endonucleolytic cleavage of chromatin characteristic of apoptosis. Most animal cells can self-destruct via an intrinsic program of cell death that may be triggered by extracellular or intracellular signals. Programmed cell death plays an important role in many biological processes such as morphogenesis and negative selection in the immune system. In addition, dysregulation of programmed cell death may contribute to diseases, for example, cancer, neurodegenerative diseases, and AIDS. Apoptosis is the term for programmed cell death characterized by specific morphologic and biochemical properties. Apoptotic cells typically undergo a series of structural changes: blebbing of the plasma membrane; condensation of the cytoplasm and nuclei; and cellular fragmentation into apoptotic bodies, membrane-bound structures containing condensed cytoplasm, some intact organelles, and nuclear fragments. The most common biochemical property of apoptosis is the endonucleolytic cleavage of chromatin, initially to large fragments of 50-300 kilobase pairs and subsequently to monomers and multimers of 180-200 base pairs. The TUNEL (Terminal deoxynucleotidyl Transferase Biotin-dUTP Nick End Labeling) method identifies apoptotic cells in situ by using terminal deoxynucleotidyl transferase (TdT) to transfer biotin-dUTP to the free 3'-OH of cleaved DNA. The biotin-labeled cleavage sites are then visualized by reaction with fluorescein conjugated avidin (avidin-FITC). Quality Control Testing: Detected cleaved DNA in HeLa cells after incubation with bovine DNase I Capacity: 40 assays using 50ul end-labeling reaction. Kit Components: T9162-80A: Proteinase K (PK), 1x400ug T9162-80B: TdT, (Terminal deoxynucleotidyl Transferase), 1x300u T9162-80C: TdT Buffer, 3x2ml T9162-80D: dUTP (Biotin), 1x100ul *T9162-80E: TB Buffer (10X), 1x50ml *T9162-80F: Blocking Solution, 1x2ml T9162-80G: Avidin (FITC), 1x10ug T9162-80H: Bovine DNase I, 1x200ug Storage and Stability: Store *T9162-80E and T9162-80F at 4 degrees C once thawed. Store other components at -20 degrees C. Stable for at least 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.