Immunoenzymatic assessment of IL-1 beta mRNA by in situ hybridization using sulphonated probes.
An immunoenzymatic detection method of in-situ hybridization reactions for interleukin 1 (IL-1) beta was established, using sulphonated probes. As model system we used unstimulated and lipopolysaccharide (LPS) stimulated peripheral blood mononuclear cells (PBMNC). After hybridization, sulphone groups were targeted with a monoclonal antibody, and bound antibody was visualized by the alkaline phosphatase anti-alkaline phosphatase (APAAP) method. In unstimulated PBMNC both the control and the IL-1 beta specific c-DNA probes were negative, whilst a large proportion of LPS treated PBMNC was positive with the sulphonated IL-1 beta plasmid only. This method may be a powerful alternative to radio-isotopic labeling. Since the entire procedure can be performed within one day, it may be applied for routine diagnostic purposes.
Gerdes
J
J
Department of Immunology and Cell Biology, Forschungsinstitut Borstel, FRG.
Herzbeck
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H
Schlüter
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Flad
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D015342:DNA Probes; D006801:Humans; D007124:Immunoenzyme Techniques; D066298:In Vitro Techniques; D007375:Interleukin-1 / Q000096:biosynthesis* / Q000235:genetics; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D013450:Sulfones
D015342:DNA Probes; D007375:Interleukin-1; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D013450:Sulfones