FD NeuroApop Kit

NDT102

NDT102 FD NeuroApopTM Kit

  • Price: $663 (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.

Detection of apoptotic neurons with FD NeuroApop™ Kit. Paraffin section (10µm) cut from a dorsal ganglion of a mouse embryo (E17). The section was processed with FD NeuroApop™ kit and counterstained with methyl green (Sections courtesy of Drs. Michael Vogel and Lisa Qiu).

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).

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 methyl green (see below)

Key Contents:

Part I (Store at -20ºC):

  • Digestive Enzyme (2 ml x 4)
  • Reaction Solution A (2 ml x 2)
  • Reaction Solution B (85 ml)
  • Reaction Solution C (60 ml)
  • Chromogen Solution (20 ml)

Part II (Store at 4ºC):

  • Equilibration Buffer (20 ml)
  • Detection Reagent (5 ml)
  • 10x Phosphate-Buffered Saline (250 ml x 2)

Materials required, but not included:

  • Double distilled water
  • Humidified chamber
  • Incubator or waterbath (30ºC)
  • 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.

Terms and Conditions

  • For quality assurance of our service, it is recommended that you discuss with us for preferred perfusion protocol and histology and/or immunolabeling protocols.
  • It is suggested that you use Gel-coated microscopic slides for tissue mounting and 0.17um-thick coverslips.
  • A 15% of the fee will be due upon authorization of the study; and the remaining fee will be due upon delivery of study results.
  • Progress of the service is contingent upon staining quality of tissues, operated by the independent contractor.
  • Should early termination occur, Neurodigitech will prorate the cost incurred and invoice the Sponsor. The first portion of the fee is non-refundable.