FD Apop™ Kit

Cat. # PK101 (for 50 sections)        Price: $429.00

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 as one of the most characteristic features of apoptosis^{2, 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 (cf. User Manual)

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:  

• 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 (PK101, Version 2000-1)

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.

References using FD Apop™ kit:

1. Thomas GR, Chen Z, Enamorado L, Bancroft C and Van Waes C: 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. International J Cancer 86:368-374, 2000.

2. Davies, JK, Shikes RH, Sze CI, Leslie A, Romero R and Gibbs RS: Histologic inflammation in the maternal and fetal compartments in a infection. Am J Obstet Gynecol. 183:1088-1093, 2000.

3. Kanamori M, Kawaguchi T, Berger MS and Pieper RO: Intracranial microenvironment reveals independent opposing functions of host αVβ3 expression on glioma growth and angiogenesis.    J. Biol. Chem. 10.1074/jbc.M605344200, 2006.                   

                                                                      (Updated 1/5/2007)