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  1. 0 資料タイプ別
  2. 01 学術雑誌論文
  1. 300 医歯学総合病院
  2. 10 学術雑誌論文
  3. 10 査読済論文

Simultaneous three-color analysis of the surface phenotype and DNA-RNA quantitation using 7-Amino-actinomycin-D and Pyronim Y

http://hdl.handle.net/10191/1168
http://hdl.handle.net/10191/1168
13a382c8-9a45-441c-a705-f8a1e83b0d35
名前 / ファイル ライセンス アクション
24_0024.pdf 24_0024.pdf (1.1 MB)
Item type 学術雑誌論文 / Journal Article(1)
公開日 2007-04-23
タイトル
言語 en
タイトル Simultaneous three-color analysis of the surface phenotype and DNA-RNA quantitation using 7-Amino-actinomycin-D and Pyronim Y
言語
言語 eng
キーワード
主題Scheme Other
主題 cell kinetics
キーワード
主題Scheme Other
主題 DNA
キーワード
主題Scheme Other
主題 RNA quantitation
キーワード
主題Scheme Other
主題 7-amino-actinomycin D
キーワード
主題Scheme Other
主題 pyronin Y
キーワード
主題Scheme Other
主題 saponin
キーワード
主題Scheme Other
主題 Flow cytometry
資源タイプ
資源 http://purl.org/coar/resource_type/c_6501
タイプ journal article
著者 Toba, Ken

× Toba, Ken

WEKO 6684

en Toba, Ken

Search repository
Winton, Elliott F.

× Winton, Elliott F.

WEKO 6685

en Winton, Elliott F.

Search repository
Koike, Tadashi

× Koike, Tadashi

WEKO 6686

en Koike, Tadashi

Search repository
Shibata, Akira

× Shibata, Akira

WEKO 6687

en Shibata, Akira

Search repository
著者別名
識別子 6688
識別子Scheme WEKO
姓名 鳥羽, 健
言語 ja
抄録
内容記述タイプ Abstract
内容記述 We developed an improved technique that permits simultaneous DNA and RNA quantitation by a flow cytofluorometry using 7-amino-actinomycin D (7AAD) and pyronin Y (PY), respectively. Detailed cell cycle analyses based upon the cellular DNA/RNA levels were performed using cells suspended in a buffer containing 0.004% saponin. This method preserved the light scattering properties of human peripheral blood cells, thus lymphocyte, monocyte and granulocyte populations could be evaluated. In addition, since 7AAD and PY exhibit red (>650 nm) and orange fluorescence (570 nm) respectively, the green fluorescence channel of the flowcytometer was reserved for surface phenotyping using FITC-conjugated antibodies. The 7AAD/PY method is applicable to the simultaneous three-color analysis of the surface phenotype and DNA-RNA quantitation when combined with FITC-conjugated surface markers in heterogeneous samples. To demonstrate the three color analysis, PHA-activated human peripheral blood lymphocytes were stained for cell surface markers with monoclonal antibodies. The cells were suspended in buffer containing 0.004% saponin, then stained with 7AAD and PY. The DNA and RNA were analyzed in indivisual CD4+, CD8+ and CD20+ cells, and the characteristic cell cycle status was found. Cell activation was further analyzed using antibodies against IL-2 receptors (CD25), transferrin receptors (CD71) or HLA-DR molecules. Transferrin receptors were expressed in late G1 phase (G1B) just before the initiation of DNA synthesis, whereas IL-2 receptors and HLA-DR were expressed very early in the G1 phase (G1T). Since this technique preserves both light scatter properties as well as cell surface proteins, it is ideally suited for detailed cell cycle analyses of heterogeneous samples such as peripheral blood or bone marrow cells.
言語 en
書誌情報 en : Journal of Immunological Methods

巻 182, 号 2, p. 193-207, 発行日 1995-02
出版者
言語 en
出版者 Elsevier
ISSN
収録物識別子タイプ ISSN
収録物識別子 0022-1759
書誌レコードID
収録物識別子タイプ NCID
収録物識別子 AA00699645
DOI
識別子タイプ DOI
関連識別子 http://doi.org/10.1016/0022-1759(95)00050-k
権利
権利情報 Elsevier B. V.
出版タイプ
出版タイプ VoR
出版タイプResource http://purl.org/coar/version/c_970fb48d4fbd8a85
著者版フラグ
値 author
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