Detection of mRNA and hnRNA using a digoxigenin labelled cDNA probe byin situ hybridization on frozen tissue sections
| Autor: | Mauro Piantelli, Arnaldo Capelli, F O Ranelletti, Riccardo Ricci, Nicola Maggiano, Luigi Maria Larocca, Libero Lauriola |
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| Rok vydání: | 1991 |
| Předmět: |
In situ
Receptors Antigen T-Cell alpha-beta T-Lymphocytes Thymus Gland In situ hybridization Biology Sensitivity and Specificity chemistry.chemical_compound Complementary DNA Frozen Sections Humans Digoxigenin RNA Messenger Frozen section procedure Infant Nucleic Acid Hybridization Cell Biology Molecular biology chemistry Cytoplasm Child Preschool RNA Heterogeneous Nuclear Alkaline phosphatase Lymph Nodes Anatomy DNA Probes Precursor mRNA |
| Zdroj: | The Histochemical Journal. 23:69-74 |
| ISSN: | 1573-6865 0018-2214 |
| DOI: | 10.1007/bf01047110 |
| Popis: | A full-length cDNA clone encoding the constant region of T cell receptor beta chain was labelled by random priming DNA with digoxigenin-dUTP. The probe was used to estimate the relative amount of the receptor beta chain mRNA by in situ hybridization on frozen sections from human thymus and lymph nodes. The hybridization was visualized in blue using an anti-digoxigenin antibody conjugated with alkaline phosphatase and a subsequent enzyme-catalysed colour reaction. The distributions of the signal in tissue sections were as expected. Moreover, labelled cells showed hybrids both in the cytoplasm and in the nucleus, and strongly and weakly stained cells were clearly distinguishable. The results indicate that this method of in situ hybridization should be useful in the detection of specific mRNA in frozen sections. |
| Databáze: | OpenAIRE |
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