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NDT101
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NDT101 FD ApopTM Kit
Price: $499.85 (For 50 sections)

This 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 as 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 identification5.

The reagents and procedure of the 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.

Contents:

Part I (Store at -20oC)
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Reaction Solution A 2 ml x 2
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Reaction Solution B 60 ul
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Reaction Solution C 40 ul
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Chromogen Solution 20 ml

Part II (Store at 4oC)
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Equilibration Buffer 20 ml
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Detection Reagent 6 ml
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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 (30oC)
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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.

References:
1.
Gavrieli Y., Sherman Y. and Ben-Sasson S. A. (1992) Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J. Cell Biol. 119: 493-501.
2.
Wyllie A. H. (1980) Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 284: 555-556.
3.
Arends M. J., Morris R. G. and Wyllie A. H. (1990) Apoptosis: the role of the endonuclease. Amer. J. Pathol. 136: 593-608.
4.
Hsu S. M., 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: 577-580.
5.

Giovana R. Thomas, Zhong Chen, Ileana Enamorado, Caren Bancroft, Carter Van Waes (2000) IL-12- and IL-2-induced tumor regression in a new murine model of oral squamous-cell carcinoma is promoted by expression of the CD80 co-stimulatory molecule and interferon-γ (2000). Int. J. Cancer: 86, 368-374.

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