Use of horseradish peroxidase- and fluorescein-modified cisplatin derivatives for simultaneous labeling of nucleic acids and proteins.
| Autor: | van Gijlswijk RP; Kreatech Biotechnology, Vlierweg 20, 1032 LG Amsterdam, The Netherlands., Talman EG, Peekel I, Bloem J, van Velzen MA, Heetebrij RJ, Tanke HJ |
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| Jazyk: | angličtina |
| Zdroj: | Clinical chemistry [Clin Chem] 2002 Aug; Vol. 48 (8), pp. 1352-9. |
| Abstrakt: | Background: Microarray platforms will change immunochemical and nucleic acid-based analysis of cell homogenates and body fluids compared with classic analyses. Microarrays use labeled target and immobilized probes, rather than fixed targets and labeled probes. We describe a method for simultaneous labeling of nucleic acids and proteins. Methods: Horseradish peroxidase- and fluorescein-modified cisplatin derivatives were used for labeling of nucleic acids and proteins. These reagents, called the Universal Linkage System (ULS), bind to sulfur- and nitrogen-donor ligands present in amino acids and nucleotides. For automated screening of proteins and nucleic acids on microarrays, it is advantageous to label these biomolecules without pre- or postpurification procedures. The labeling of antibodies and nucleic acids in whole serum was therefore pursued. Results: Immunoglobulins in nonpurified serum were labeled efficiently enough to be used for immunochemistry. To investigate whether protein-adapted labeling allowed nucleic acid labeling as well, 1 microg of plasmid DNA was added to 1 microL of serum. DNA and serum proteins were simultaneously labeled, and this labeled DNA could be used as a probe for direct fluorescence in situ hybridization. Conclusion: ULS provides a direct labeling tool for the (simultaneous) modification of proteins and nucleic acids even in unpurified samples. |
| Databáze: | MEDLINE |
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