Quantitative immunofluorescence assay for cyclobutyldithymidine dimers in individual mammalian cells

Lesko, Stephen A.; Li, W.; Zheng, Gang; Callahan, D.E.; Kaplan, David S.; Midden, W. Robert; Strickland, Paul T.

doi:10.1093/carcin/10.4.641

Cited by 17 publications

(4 citation statements)

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“…WBCs were fiied in methanokacetic acid (91) at room temperature (RT) and stored at -20'C. Before analysis, WBCs were put on multiwell slides (Kirkegaard & Perry) with a pipet and were processed as described by L.esko et al (21). with slight modifications.…”

Section: Methodsmentioning

confidence: 99%

Detection of DNA adducts in basal and non-basal cells of the hamster trachea exposed to benzo(a)pyrene in organ culture.

Roggeband¹,

Berg²,

Wulp³

et al. 1994

J Histochem Cytochem.

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DNA adducts were quantified in hamster tracheas exposed to benzo(a)pyrene (BP) in organ culture, in basal as well as in non-basal cells, by in situ detection with an adduct-specific rabbit antiserum (W2/01) and with a mouse monoclonal antibody against human cytokeratins 5 and 8 (RCK102) to identify hamster trachea basal cells. Recognition by W2/01 of the adduct of (+)-anti-7,8-dihydmxy-9,lO-epoxide of BP (BP diolepoxide; BPDE) to deoxyguanosine (dG) was checked on human white blood ells (WBCs) exposed to BP together with 3-methylcholanthrene (3MC)-induced rat-liver miaosomes. By comparison with the adduct levels determined by 32P post-labeling, a lower detection limit of about 1 adduct per lo6 nucleotides could be deduced. Next, tracheal rings were exposed to BP (40 pM) in organ culture for 2 days, then

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Section: Methodsmentioning

confidence: 99%

Detection of DNA adducts in basal and non-basal cells of the hamster trachea exposed to benzo(a)pyrene in organ culture.

Roggeband¹,

Berg²,

Wulp³

et al. 1994

J Histochem Cytochem.

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“…In 1983 immunofluorescence microscopy (IFM) was used to visualize points of UV damage induced in anaphase nuclei produced by laser microirradiation and the subsequent restricted diffusion of the damaged regions as the cell entered interphase (43). Over the next few years IHC and microfluorometry were applied to (6‐4) PD analyses in cells (44–47) and skin (48–50). Similar procedures were developed using CPD MAbs, including IFM studies to detect a visible light enhancement of CPD removal in human epidermis treated with a multiple split‐dose (but not single dose) UVR (51) and laser scanning confocal microscopy to construct a three‐dimensional image of the nuclear localization of CPDs, (6‐4) PDs and unscheduled DNA synthesis (52).…”

Section: Technology Developmentmentioning

confidence: 99%

Antibodies and DNA Photoproducts: Applications, Milestones and Reference Guide

Mitchell¹,

Brooks

2010

Photochem & Photobiology

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Polyclonal and monoclonal antibodies recognizing the primary DNA photoproducts induced by ultraviolet radiation (UVR) have proven to be essential tools in the study of photochemical and photobiological phenomena. As specific ''DNA damage binding proteins'' these reagents have been used to develop a diverse array of technical procedures applied to a plethora of important problems in DNA photochemistry and the biological effects of UVR at the molecular, developmental, organism and population levels. This survey attempts to cover this science from an historical perspective and to reveal the great breadth of discovery and contribution associated with the development and application of DNA damage antibodies to the current body of science.Three papers published in the 1960s were seminal to the development of immunoassays for the study of photochemical and photobiological phenomena. In 1960 Rosalyn Yalow and Solomon Berson published a paper in the Journal of Clinical Investigation entitled ''Immunoassay of endogenous plasma insulin in man'' (1), which described the development of a sensitive radioimmunoassay (RIA) to measure minute quantities of hormones in blood samples. This technique played a seminal role in the exponential expansion of research in medicine and the biological sciences over the past half century. In 1977 Dr. Yalow was awarded the Nobel Prize in Medicine for her work. In 1964 Otto Plescia and colleagues published a paper in the Proceedings of the National Academy of Sciences entitled ''Production of antibodies to denatured deoxyribonucleic acid (DNA)'' (2). Using a technique whereby singlestranded DNA was electrostatically coupled to methylated bovine serum albumin they were able to elicit an immune response in rabbits. In the wake of this paper it was not long before a group led by Lawrence Levine and Helen van Vanukis published an article in Science in 1966 entitled ''Antibodies to photoproducts of deoxyribonucleic acids irradiated with ultraviolet light'' (3). Using the technique established by Plescia, this group was able to produce a rabbit antiserum using DNA irradiated with 270 nm light with antigenic determinants that were reversible by 235 nm light and responsive to UV-irradiated thymidine nucleotides. It appeared that antibodies had been created in response to thymine dimers.Over the course of the following two decades a number of polyclonal and monoclonal antibodies to various UV-induced DNA lesions were produced and characterized. The technology that developed blossomed and has been applied to a broad spectrum of problems in photochemistry, the biological response to UV radiation (UVR), photomedicine and environmental photobiology. Early immunoassays generated DNA repair curves showing rapid removal of most antibody binding sites leaving behind a significant remnant (4,5). These results were not consistent with the known repair kinetics using biochemical and enzymatic techniques. Subsequently, it was discovered that polyclonal antisera raised against heavily UVirradiated DNA contai...

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“…Several analytical approaches, including 32 P-postlabeling assays ( − ), immunological methods ( , ), HPLC in combination with electrochemical detection, gas chromatography coupled to mass spectrometry ( , ), and capillary electrophoresis ( , ), have been developed for the detection and the measurement of DNA base modifications (for comprehensive reviews, see refs 14 and 15 ).…”

Section: Introductionmentioning

confidence: 99%

Polyclonal Antibodies to Adenine N¹-Oxide: Characterization and Use for the Measurement of DNA Damage

Signorini

Molko²,

Cadet³

1998

Chem. Res. Toxicol.

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Adenine N1-oxide is a DNA lesion whose formation involves the specific oxidation of the adenine base by hydrogen peroxide under nonradical conditions. The damage may be measured using a HPLC/32P-postlabeling method, which however cannot be used for routine analysis. We propose herein as an alternative an immunological assay which allows a rapid evaluation of the level of adenine N1-oxide in DNA exposed to oxidative stress. Two polyclonal antibodies were raised using two different strategies for the coupling of the hapten to the protein. The first approach is based on the universal method of Erlanger and Beiser, whereas the preparation of the second antigen involves the conjugation of a morpholino derivative of adenosine N1-oxide to the carrier protein. The affinity and the specificity of those antibodies were determined by competitive enzyme-linked immunosorbent assay. The antibody obtained by the traditional method shows some cross-reactivity with normal nucleotides, whereas for the other antiserum, the selectivity was found to be higher. Therefore, this polyclonal antibody was used to quantify the level of adenine N1-oxide in calf thymus DNA oxidized either by m-chloroperbenzoic acid or by hydrogen peroxide. The detection limit of the assay is four residues of adenine N1-oxide per 10(6) normal bases. The level of adenine N1-oxide in nonmodified DNA was lower than the detection limit of the assay, whereas in mCPB- and H2O2-modified DNA, it could be up to 14 and 0.7 adenine N1-oxide residues per 10(4) normal bases, respectively.

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Quantitative immunofluorescence assay for cyclobutyldithymidine dimers in individual mammalian cells

Cited by 17 publications

References 0 publications

Detection of DNA adducts in basal and non-basal cells of the hamster trachea exposed to benzo(a)pyrene in organ culture.

Detection of DNA adducts in basal and non-basal cells of the hamster trachea exposed to benzo(a)pyrene in organ culture.

Antibodies and DNA Photoproducts: Applications, Milestones and Reference Guide

Polyclonal Antibodies to Adenine N¹-Oxide: Characterization and Use for the Measurement of DNA Damage

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Quantitative immunofluorescence assay for cyclobutyldithymidine dimers in individual mammalian cells

Cited by 17 publications

References 0 publications

Detection of DNA adducts in basal and non-basal cells of the hamster trachea exposed to benzo(a)pyrene in organ culture.

Detection of DNA adducts in basal and non-basal cells of the hamster trachea exposed to benzo(a)pyrene in organ culture.

Antibodies and DNA Photoproducts: Applications, Milestones and Reference Guide

Polyclonal Antibodies to Adenine N1-Oxide: Characterization and Use for the Measurement of DNA Damage

Contact Info

Product

Resources

About

Polyclonal Antibodies to Adenine N¹-Oxide: Characterization and Use for the Measurement of DNA Damage