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Quantitation of 5-bromo-2-deoxyuridine incorporation into DNA: an enzyme immunoassay for the assessment of the lymphoid cell proliferative response |
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| Authors: | T Porstmann T Ternynck S Avrameas |
| Affiliation: | 1. Institute of Medical Immunology, Faculty of Medicine (Charité), Humbold University of Berlin, Schumannstrasse 20/21, 1040 Berlin, G.D.R.;2. Unité d''Immunocytochimie, Département d''Immunologie, Institut Pasteur, 25, rue du Docteur Roux, 75724 Paris Cedex 15, France |
| Abstract: | As an alternative to the measurement of radiolabeled thymidine incorporated into DNA, a method is presented in which thymidine has been replaced by its analogue, 5-bromo-2-deoxyuridine (BUdR). BUdR incorporated into DNA (BUdR-DNA) is measured by a sandwich-type enzyme immunoassay using a monoclonal anti-BUdR antibody. This method allows the quantitation of 4 ng of BUdR-DNA. Comparative experiments with myeloma cells and LPS stimulated spleen B-cells have shown that this technique is at least as sensitive as the traditional counting of [3H]thymidine. |
| Keywords: | monoclonal anti-BUdR antibody 5-bromo-2-deoxyuridine (BUdR) incorporation into DNA DNA extraction enzyme immunoassay of BUdR immunocytochemical staining DNA deoxyribonucleic acid BUdR 5-bromo-2-deoxyuridine Thy thymidine BUdR-DNA BUdR in vivo labeled DNA BSA bovine serum albumin OVA ovalbumin BU 5-bromouridine BU-BSA (or BU-OVA) BU conjugated to BSA (or OVA) LPS lipopolysaccharide mAb monoclonal antibody HRP horseradish peroxidase SDS sodium dodecyl sulfate EIA enzyme immunoassay EDTA ethylenediaminetetraacetic acid PBS 0.15 M NaCl, 0.01 M phosphate buffer pH 7.5 TE buffer 0.01 Tris-HCl buffer pH 7.5, 0.1 M EDTA X63 X63-Ag 8.653 myeloma cells |
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