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NDT102
NDT101 NDT103 NDT104 NDT105  

Detection of apoptotic neurons with NDT102 Kit. Paraffin section (10um) cut from a dorsal ganglion of a mouse embryo (E17). The section was processed with the kit and counterstained with methyl green (Sections courtesy of Drs. Michael Vogel and Lisa Qiu)
(click to see enlarged photo)
Detection of apoptotic neurons with NDT102 Kit. Paraffin section (10um) cut from a dorsal ganglion of a mouse embryo (E17). The section was processed with the kit and counterstained with methyl green (Sections courtesy of Drs. Michael Vogel and Lisa Qiu)
(click to see enlarged photo)
Detection of apoptotic neurons in a rat model of stroke. 20 um cryostat section was cut from the rat striatum of a stroke model. The section was processed for detecting neuronal apoptosis with the kit and then counterstained with NDT methyl green (see below)
(click to see enlarged photo)
NDT102 FD NeuroApopTM Kit
Price: $586.30 (For 35 sections)

This Kit is specifically designed for the detection of neuronal apoptosis in tissue sections from the central nervous system 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 identification.

The reagents and procedure of the kit have been optimized to achieve a high degree of both specificity and sensitivity for detecting apoptotic neurons with the lowest background. This kit can be used with frozen5 and paraffin sections, as well as cultured cells (cf. photo samples below). The procedure of the kit takes approximately 4 hours.

Contents:

Part I (Store at -20ºC)
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Digestive Enzyme 2 ml x 4
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Reaction Solution A 2 ml x 2
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Reaction Solution B 85 ml
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Reaction Solution C 60 ml
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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 5 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.
Wei H, Qin Z.-H., Senatorov V. V., Wei W., Wang Y., Qian Y. and Chuang D.-M. (2001) Lithium suppresses excitotoxicity-induced striatal lesions in a rat model of Huntington's disease. Neuroscience 106: 603-612.

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