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An antibody against single-stranded DNA useful for detecting apoptotic cells : production and applications for development, differentiation and carcinogenesis

Research Project

Project/Area Number 08557013
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Pathological medical chemistry
Research InstitutionAkita University

Principal Investigator

SUGIYAMA Toshihiro  Akita University School of Medicine Professor, 医学部, 教授 (00127242)

Co-Investigator(Kenkyū-buntansha) URAYAMA Osamu  Akita University School of Medicine Associate Professor, 医学部, 助教授 (90114743)
TAKADA Goro  Akita University School of Medicine Professor, 医学部, 教授 (40047254)
TANAKA Toshinobu  Akita University School of Medicine Professor, 医学部, 教授 (40002216)
KOYAMA Kenji  Akita University School of Medicine Professor, 医学部, 教授 (80004638)
TOMITA Yasushi  Nagoya University School of medicine Professor, 医学部, 教授 (70108512)
Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥12,700,000 (Direct Cost: ¥12,700,000)
Fiscal Year 1997: ¥5,500,000 (Direct Cost: ¥5,500,000)
Fiscal Year 1996: ¥7,200,000 (Direct Cost: ¥7,200,000)
Keywordsapoptosis / ssDNA / single-stranded DNA / differentiation / development / antibody / carcinogenesis / 発生・分化 / 単離DNA
Research Abstract

In this research project we developed an improved method for the production of the antibody and investigated into the antigenic determinants of the antibody so that it could be of practical use for detecting apoptotic cells. Rabbits, hyperimmunized with complexes of alkaline-denatured calf thymus DNA and methylated bovine serum albumin, produced an IgG antibody to single-stranded DNA.Analysis by sandwich ELISA using various naturally occurring nucleic acids revealed that the antibody was specific to single-stranded DNA.Furthermore, using synthetic polymers in the assay, it was found that the antibody could recognize single-stranded DNA with various base sequences. Gel electrophoresis retardation assays, with synthetic oligodeoxynucleotides with differing lengths of single-stranded DNA,indicated that a hexadeoxynucleotide constituted the minimum size of the antigenic determinants, and suggested that the antibody probably consists of several antibodies which recognize hexadeoxynucleotides with various base sequences. Western blot analysis demonstrated that the antibody can recognize both a DNA ladder and oligonucleosomes prepared from rat liver nuclei with endogenous endonuclease. We also demonstrated that this antibody is a useful tool for detecting apoptotic cells in development, differentiation and carcinogenesis.

Report

(3 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • Research Products

    (20 results)

All Other

All Publications (20 results)

  • [Publications] Kawarada, Y.: "Antibody against single-stranded DNA useful for detecting apoptotic cells recognizes hexadeoxynucleotides with various base sequences." J.Biochem.123. 492-498 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Maeda, M.: "Single-stranded DNA as an immunocytochemical marker for apoptotic change of ishemia in the gerbil hippocampus." Neurosci.Lett.240. 69-72 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Umemura, S.: "Enhancement of TdT-mediated dUTP-biotin nick end labeling(TUNEL)method using mung bean nuclease,a single-stranded DNA digesion enzyme." J Histochem.Cytochem.44. 125-132 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 張 鳳春: "LECラット(CD4細胞欠乏ラット)胸腺細胞におけるアポトーシスの研究" 秋田医学. 24. 51-58 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Maeda, M: "Single stranded DNA as an immunocytochemical marker for apoptotic change of ischemia in the gerbil hippocampus." Neurosci.Lett.(in press). (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kawarada, Y.: "Antibody against single-stranded DNA useful for detecting apoptotic cells recognizes hexadeoxynucleotides with various base sequences." J.Biochem.(in press). (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Umemura, S.: "Enhancement of TdT-mediated dUTP-biotin nick end labeling (TUNEL) method using mung bean nuclease, a single-stranded DNA digestion enzyme." J.Histochem.Cytochem.44. 125-132 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Zhang, F.: "Study of apoptosis in LEC rat thymocyte of CD4 cell deficiency" Akita J.Med.24. 51-58 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kawarada,Y: "Antibody against single-stranded DNA useful for detecting apoptotic cells recognizes hexadeoxynucleotides with various base sequences." J.Biochem.(in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Maeda,M.: "Single-stranded DNA as an immunocytochemical marker for apoptotic change of ishemia in the gerbil hippocampus." Neurosci.Lett.(in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Terada,K.: "Restoration of holoceruloplasmin synthesis in Lec rat after infusion of recombinant adenovirus bearing WND cDNA." J.Biol.Chem.273. 1815-1820 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Yasui,O.: "Isolation of oval cells from Long-Evans Cinnamon rats and their transformation into hepatocytes in vivo in the rat liver." Hepatology. 25. 329-334 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Yang,X-L.: "Two forms of Wilson disease proteins produced by alternative splicing are localized in destinct cellular compartments." Biochem.J.326. 897-902 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 張 鳳春: "LECラット(CD4細胞欠乏ラット)胸腺細胞におけるアポトーシスの研究" 秋田医学. 24. 51-58 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Yasui,O.: "Isolation of oval cells from Long-Evance Cinnamon rats and their transformation into hepatocytes in vivo in the rat liver." Hepatology. 25. 329-334 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Masuda,Y.: "Platelet-derived growth factor B-chain homodimer suppressing a convulsion of epilepsy model mouse El." Biochem.Biophys.Res.Commun.223. 60-63 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Umemura,S.: "Enhancement of TdT-mediated dUTP-biotin nick end labeling (TUNEL) method using mung bean nuclease,a single-stranded DNA digestion enzyme." J.Histochim.Cytochem.44. 125-132 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hirasawa,F.: "The effect of silver administration on the biosynthesis and the molecular properties of rat cerulopasmin." Biochim.Biophys.Acta. (in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Yamazaki,Y.: "The use of capillary liquid chromatography/electrospray mass spectrometry to identify a specific systeine residue susceptible to S-thiolation in recombinant human Cu,Zn superoxide dismutase." Res.Commun.Biochem.Cell.Biol.(in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kuhara,M.: "Enhancing effect of a congenital disorder in methionine metabolism on the development of spontaneous hepatitis and hepatoma in LEC rats." J.Trace Elem.Exp.Med.(in press). (1997)

    • Related Report
      1996 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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