Immunochemical detection of DNA in alkaline sucrose gradient fractions

J. Vijg; E. Mullaart; L. Roza; R. A. Baan; P. H.M. Lohman

doi:10.1016/0022-1759(86)90101-8

Immunochemical detection of DNA in alkaline sucrose gradient fractions

J. Vijg, E. Mullaart, L. Roza, R. A. Baan, P. H.M. Lohman

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

A procedure for the detection of non-radioactive DNA in alkaline sucrose gradients is described. This method consists of the following steps: (1) fractionation of the DNA-containing gradients into microtiter plates, neutralization and overnight adsorption; (2) covalent labelling of the guanine bases of the adsorbed DNA by reaction with N-acetoxy-2-acetylaminofluorene; and (3) determination of the gradient profile by means of an enzyme-linked immunosorbent assay using antibodies with high affinity for dG-AAF. The method has been found suitable for the rapid and sensitive determination of gamma-ray induced single-strand breaks and UV-induced pyrimidine dimers in mammalian cells in vitro. In addition, it has been shown that the method can be used for the determination of pyrimidine dimers induced in skin cells in vivo.

Original language	English (US)
Pages (from-to)	53-58
Number of pages	6
Journal	Journal of Immunological Methods
Volume	91
Issue number	1
DOIs	https://doi.org/10.1016/0022-1759(86)90101-8
State	Published - Jul 11 1986
Externally published	Yes

Keywords

Alkaline sucrose gradient
Automated assay
DNA
DNA repair
Immunochemical method
Pyrimidine dimer

ASJC Scopus subject areas

Immunology and Allergy
Immunology

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10.1016/0022-1759(86)90101-8

Cite this

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title = "Immunochemical detection of DNA in alkaline sucrose gradient fractions",

abstract = "A procedure for the detection of non-radioactive DNA in alkaline sucrose gradients is described. This method consists of the following steps: (1) fractionation of the DNA-containing gradients into microtiter plates, neutralization and overnight adsorption; (2) covalent labelling of the guanine bases of the adsorbed DNA by reaction with N-acetoxy-2-acetylaminofluorene; and (3) determination of the gradient profile by means of an enzyme-linked immunosorbent assay using antibodies with high affinity for dG-AAF. The method has been found suitable for the rapid and sensitive determination of gamma-ray induced single-strand breaks and UV-induced pyrimidine dimers in mammalian cells in vitro. In addition, it has been shown that the method can be used for the determination of pyrimidine dimers induced in skin cells in vivo.",

keywords = "Alkaline sucrose gradient, Automated assay, DNA, DNA repair, Immunochemical method, Pyrimidine dimer",

author = "J. Vijg and E. Mullaart and L. Roza and Baan, {R. A.} and Lohman, {P. H.M.}",

year = "1986",

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doi = "10.1016/0022-1759(86)90101-8",

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T1 - Immunochemical detection of DNA in alkaline sucrose gradient fractions

AU - Vijg, J.

AU - Mullaart, E.

AU - Roza, L.

AU - Baan, R. A.

AU - Lohman, P. H.M.

PY - 1986/7/11

Y1 - 1986/7/11

N2 - A procedure for the detection of non-radioactive DNA in alkaline sucrose gradients is described. This method consists of the following steps: (1) fractionation of the DNA-containing gradients into microtiter plates, neutralization and overnight adsorption; (2) covalent labelling of the guanine bases of the adsorbed DNA by reaction with N-acetoxy-2-acetylaminofluorene; and (3) determination of the gradient profile by means of an enzyme-linked immunosorbent assay using antibodies with high affinity for dG-AAF. The method has been found suitable for the rapid and sensitive determination of gamma-ray induced single-strand breaks and UV-induced pyrimidine dimers in mammalian cells in vitro. In addition, it has been shown that the method can be used for the determination of pyrimidine dimers induced in skin cells in vivo.

AB - A procedure for the detection of non-radioactive DNA in alkaline sucrose gradients is described. This method consists of the following steps: (1) fractionation of the DNA-containing gradients into microtiter plates, neutralization and overnight adsorption; (2) covalent labelling of the guanine bases of the adsorbed DNA by reaction with N-acetoxy-2-acetylaminofluorene; and (3) determination of the gradient profile by means of an enzyme-linked immunosorbent assay using antibodies with high affinity for dG-AAF. The method has been found suitable for the rapid and sensitive determination of gamma-ray induced single-strand breaks and UV-induced pyrimidine dimers in mammalian cells in vitro. In addition, it has been shown that the method can be used for the determination of pyrimidine dimers induced in skin cells in vivo.

KW - Alkaline sucrose gradient

KW - Automated assay

KW - DNA

KW - DNA repair

KW - Immunochemical method

KW - Pyrimidine dimer

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JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

IS - 1

ER -

Immunochemical detection of DNA in alkaline sucrose gradient fractions

Abstract

Keywords

ASJC Scopus subject areas

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