FD Apop™ Kit
FD Apop™ Kit is designed for the microscopic detection of cells undergoing apoptosis based on the principle of in situ DNA nick-end labeling (TUNEL) technique¹. The assay uses terminal deoxynucleotidyl transferase to catalyze the incorporation of biotinylated deoxyuridines onto the free 3′-hydroxyl termini of DNA fragments, which are considered one of the most characteristic features of apoptosis2, 3. The integrated biotins are amplified and visualized with the avidin-biotin-complex (ABC) method4, enabling light microscopic identification.
The reagents and procedure of FD Apop™ Kit have been optimized to achieve a high degree of both specificity and sensitivity for detecting apoptotic cells with minimal background. This kit can be used with frozen and paraffin sections, as well as cultured cells. The procedure of the kit takes approximately 4 hours.
Kit Contents
Part I (Store at -20°C)
Digestive Enzyme 2 ml x 4 Reaction Solution A 2 ml x 2 Reaction Solution B 60 µl Reaction Solution C 40 µl Chromogen Solution 20 ml Part II (Store at 4°C)
Equilibration Buffer 20 ml Detection Reagent 6 ml 10x Phosphate-Buffered Saline 250 ml x 2 Materials Required But Not Included
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Double distilled water
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Humidified chamber
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Incubator or waterbath (30°C)
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Histological supplies and equipment, including microscope slides, glass coverslips, staining jars, fine-tipped forceps, ethanol, xylenes or xylene-substitutes, mounting medium, and a light microscope.
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References
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Gavrieli Y, Sherman Y, and Ben-Sasson SA. (1992) Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J. Cell Biol. 119:493-501.
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Wyllie AH. (1980) Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature. 284:555-6.
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Arends MJ, Morris RG, and Wyllie AH. (1990) Apoptosis: the role of the endonuclease. Amer. J. Pathol. 136:593-608.
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Hsu SM, Raine L, and Fanger H. (1981) Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J. Histochem. Cytochem. 29:577-80.
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