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Immunological staining of BrdUrd : evaluation of DNA denaturation using DNase

山口医学 Volume 38 Issue 2 Page 163-169
published_at 1989-04
Title
免疫組織化学的 BrdUrd 染色 : DNase による DNA 変性法の検討
Immunological staining of BrdUrd : evaluation of DNA denaturation using DNase
Creators Matsumura Kouji
Creators Sasaki Kohsuke
Creators Tsuji Tatsuo
Creators Murakami Tomoyuki
Creators Takahashi Manabu
Creators Shinozaki Fumihiko
Source Identifiers
In cell kinetic research, there is an increasing interest in the use of immunohistochmistry for the detection of bromo-deoxyuridine (BrdUrd), which S-phase cells have incorporated into their DNA. In order to make the incorporated BrdUrd accessible to an anti-BrdUrd antibody (B-44), DNA denaturation is required. Usually this has been accomlished by exposing the cells to acid. However, the procedure using acid may destory cellular membrane antigens as well as morphological details. BU-1 antibody, when used in combination with DNase for DNA denaturation, reacts with the incorporated BrdUrd without using acid. In this study, the efficacy of BU-1 antibody was investigated and compared with that of B-44 antibody. Ten tumor speciments were used for this study. Under varying conditions (one or three atomospheric pressure during the BrdUrd lebeling and different temperture during the incubation of BU-1 antibody), the tumor tissue was labeled with BrdUrd in vitro and the each sections were immunohistochemically stained with BU-1 antibody or B-44 antibody. Three atmosppheric pressure was required for labeling cells with BrdUrd in vitro. The stainability of BrdUrd was enhanced, when tissue specimens were incubated at 37℃. The BU-1 antibody provided results equivalent to the B-44 antibody. Double staining of BrdUrd and epithelial membrane antigen (EMA) was carried out. EMA and cellular morphology were better preserved in preparations stained with BU-1 antibody than with B-44 antibody.
Subjects
医学 ( Other)
Languages jpn
Resource Type journal article
Publishers 山口大学医学会
Date Issued 1989-04
File Version Not Applicable (or Unknown)
Access Rights metadata only access
Relations
[ISSN]0513-1731
[NCID]AN00243156
Schools 医学部