Unit Code 80785:
Kappa and Lambda Light Chain Messenger RNA Detection by In Situ Hybridization
Method Description
Routinely fixed, paraffin-embedded tissue sections are deparaffinized
and digested with proteinase. Fluorescein isothiocyanate (FITC)-
conjugated kappa and lambda mRNA-specific oligonucleotides and
negative control are added to the tissue sections. After overnight
hybridization, FITC-AP conjugate is allowed to react with the fluorescein
groups on the probe molecules. Finally, the sight of hybridization is
visualized by the colorimetric reaction of the enzymes conjugate with its
substrate, BCIP (5-bromo-4-chloro-3-indolyl phosphate), and the
concomitant reduction of nitro blue tetrazolium. This reaction results in the
deposition of an insoluble blue-purple product at the site of hybridization.
(Pringle JH, Ruprai AK, Primrose L, et al: In-situ hybridization of
immunoglobulin light chain mRNA in paraffin sections using biotinylated or
hapten-labelled oligonucleotide probes. J Pathol 1990;162:197-207;
Package insert: Dako Kappa/Lambda mRNA ISH kit, Dako Corporation,
Carpinteria, CA)
Performing Laboratory Location
Rochester
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