Chapter 2
In Situ Nick Translation
SATOSHI HASHIMOTO AND TAKASHI KANEMATSU
Introduction
Nick (DNA single strand break) have been implicated to play
important roles in the regulation of cell proliferation (Ogawa
1980), cell differentiation (Appleby 1977) and cell death (Peitsch
1993). Nick translation detects nicks in DNA by adding nucleotides to 3' -hydroxyl ends in the presence of a template, extending
the strand in the 5' to 3' direction. (Fig. 1) In situ nick translation
(ISNT)(Dawson 1980) is application of nick translation on frozen sections or formalin fixed paraffin embedded tissues.
Outline
1. Preparation of samples
2. Nick translation reaction
3. Blocking and immunohistochemistry
4. Visualization
Materials
- 3-aminopropyltriethoxysilane (Sigma, USA) coated slides
Correspondence to Satoshi Hashimoto, Nagasaki University School ofMedicine, Second Department of Surgery, 1-7-1, Sakamoto, Nagasaki, 852,
Japan (phone +81-95-849-7316; fax +81-95-849-7319; e-mail hashis@mb.infoweb.ne.jp); Takashi Kanematsu, Nagasaki University School of Medicine, Second Department of Surgery, 1-7-1, Sakamoto, Nagasaki, 852, Japan
In Situ Nick Translation
SATOSHI HASHIMOTO AND TAKASHI KANEMATSU
Introduction
Nick (DNA single strand break) have been implicated to play
important roles in the regulation of cell proliferation (Ogawa
1980), cell differentiation (Appleby 1977) and cell death (Peitsch
1993). Nick translation detects nicks in DNA by adding nucleotides to 3' -hydroxyl ends in the presence of a template, extending
the strand in the 5' to 3' direction. (Fig. 1) In situ nick translation
(ISNT)(Dawson 1980) is application of nick translation on frozen sections or formalin fixed paraffin embedded tissues.
Outline
1. Preparation of samples
2. Nick translation reaction
3. Blocking and immunohistochemistry
4. Visualization
Materials
- 3-aminopropyltriethoxysilane (Sigma, USA) coated slides
Correspondence to Satoshi Hashimoto, Nagasaki University School ofMedicine, Second Department of Surgery, 1-7-1, Sakamoto, Nagasaki, 852,
Japan (phone +81-95-849-7316; fax +81-95-849-7319; e-mail hashis@mb.infoweb.ne.jp); Takashi Kanematsu, Nagasaki University School of Medicine, Second Department of Surgery, 1-7-1, Sakamoto, Nagasaki, 852, Japan