FD NeuroApop™ Kit

Cat. # PK201 (for 35 sections)       Price: $456.50 

FD NeuroApop™ 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) techniqueą. 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 FD NeuroApop™ 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 frozen and paraffin sections, as well as cultured cells (cf. below for photo samples and references using this kit).  The procedure of the kit takes approximately 4 hours (cf. User Manual).


           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 (click to see enlarged photo). 

Detection of apoptotic neurons with FD NeuroApop™ kit. Paraffin section (10µm) cut from the inferior colliculus 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 (click to see enlarged photo). 

Detection of apoptotic neurons in a rat model of stroke.  20 µm cryostat section was cut from the rat striatum of a stroke model. The section was processed for detecting neuronal apoptosis with FD NeuroApop™ Kit  and then counterstained with FD methyl green (click to see enlarged photo).

 

Kit Contents: 

Part I (Store at -20°C) 

  • Digestive Enzyme                                    2 ml x 4

  • Reaction Solution A                                 2 ml x 2

  • Reaction Solution B                                 85 µl

  • Reaction Solution C                                 60 µl

  • Chromogen Solution                                20 ml

Part II (Store at 4°C)  

  • 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 (30°C)

  • 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.  

User Manual (PK201, Version 2002-1)

 

References:

  1. Gavrieli Y, Sherman Y and Ben-Sasson SA, Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J. Cell Biol. 119:493-501, 1992.

  2. Wyllie AH. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 284:555-556, 1980.

  3. Arends MJ, Morris RG and Wyllie AH. Apoptosis: the role of the endonuclease. Amer. J. Pathol. 136: 593-608, 1990.  

  4. Hsu SM, Raine L and Fanger H. 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, 1981.  

References using FD NeuroApop™ kit:

  1. Liang ZQ, Li YL, Zhao XL, Han R, Wang XX, Wang Y, Chase TN, Bennett  MC and Qin ZH. NF-κB contributes to 6-hydroxydopamine-induced apoptosis of nigral dopaminergic neurons through p53.  Brain Research   1145:190-203, 2007.

  2. de Rivero Vaccari JC, Casey GP, Aleem S, Park WM and Corriveau RA. NMDA receptors promote survival in somatosensory relay nuclei by inhibiting Bax-dependent developmental cell death.  Proc. Natl. Acad. Sci. USA 103:16971-16976, 2006.

  3. Williams AJ, Hartings JA, Lu  XCM, Rolli ML and Tortella FC. Penetrating ballistic-like brain injury in the rat: differential time courses of hemorrhage, cell death, inflammation, and remote degeneration. J. Neurotrauma 23:1828-1846, 2006.

  4. Garris DR. Cytochemical analysis of Pancreatic islet lipoapoptosis: hyperlipidemia-induced cytoinvolution following expression of the diabetes (db/db) mutation. Pathobiology 72:124-132, 2005.

  5. Garris DR. Diabetes (db/db) mutation-induced endometrial epithelial lipoapoptosis: ultrastructural and cytochemical analysis of reproductive tract atrophy. Reproductive Biology and Endocrinology 3:15, 2005.

  6. Garris DR. Gonadal steroid modulation of the diabetes (db/db) mutation-induced hyperlipometabolic, hypogonadal syndrome: Restoration of female reproductive tract cytochemical and structural indices.  Pathophysiology  12:109-120, 2005.

  7. Lau YS, Novikova L and Roels C. MPTP treatment in mice does not transmit and cause Parkinsonian neurotoxicity in non-treated cagemates through close contact.  Neuroscience Research   52:371-378, 2005.

  8. Lei L, Laub F, Lush M, Romero M, Zhou J, Luikart B, Klesse L, Ramirez F and Parada LF. The zinc finger transcription factor Klf7 is requited for TrkA gene expression and development of nociceptive sensory neurons. Genes & Development 19:1354-1364, 2005.

  9. Cernak I, Vink R, Zapple DN, Cruz MI, Ahmed F, Chang T, Fricke ST and Faden AI. The pathobiology of moderate diffuse traumatic brain injury as identified using a new experimental model of injury in rats. Neurobiol. Dis. 17:29-43, 2004.

  10. Garris BL, Novikova L, Lau Y-S, Garris DR. Hypophyseal lipoapoptosis: diabetes (db/db) mutation-associated cytolipidemia promotes pituitary cellular disruption and dysfunction. Pituitary 7:5-14, 2004.

  11. Garris DR, Novikova L, Garris BL, Lau YS. Hypercytolipidemia-induced nuclear lipoapoptosis: cytochemical analysis and integrated review of hypogonadal, diabetes-obesity syndrome-induced female reproductive axis disruption.  Metab Syndr & Related Dis.  2:198-209, 2004.

  12. Saito R,  Bringas JR, Panner A, Tamas M, Pieper RO, Berger MS and Bankiewicz KS. Convection-enhanced delivery of tumor necrosis factor-related apoptosis-inducing ligand with systemic administration of temozolomide prolongs survival in an intracranial glioblastoma xenograft model. Cancer Res. 64:6858-6862, 2004.

  13. Wei H, Qin ZH, Senatorov VV, Wei W, Wang Y, Qian Y and Chuang DM.  Lithium suppresses excitotoxicity-induced striatal lesions in a rat model of Huntington's disease. Neuroscience 106:603-612, 2001.

  14. Davies JK, Shikes RH, Sze CI, Leslie KK, McDuffie RS, Romero R and Gibbs RS: Histologic inflammation in the maternal and fetal compartments in a rabbit model of acute intra-amniotic infection. Am. J. Obstet Gynecol. 183:1088-1093, 2000.

                                                                               (Updated 7/13/2007)

                                 Home  | Services | Products | Consulting | Order Info | Policies | Contact Us