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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
medicine
In situ hybridization of immunoglobulin light chain mRNA in paraffin sections using biotinylated or hapten‐labelled oligonucleotide probes
The Journal of Pathology, Volume 162, No. 3, Year 1990
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Description
An in situ hybridization technique has been developed for the detection of immunoglobulin light chain mRNA in routine pathology specimens. The method detects k or λ constant region sequences using a cocktail of synthetic oligonucleotide probes labelled with biotin or fluorescein 5‐isothiocyanate (FITC) reporter molecules. The probes were labelled at flanking sites chemically by primary amine directed acylation and by ‘homopolymer tailing’ with terminal deoxynucleotidyl transferase using non‐radioactive nucleotide analogues. The mRNA was unmasked in the formalin‐fixed tissue sections by digestion with varying concentrations of proteinase K, and the hybrids were demonstrated using alkaline phosphatase with either a streptavidin/biotin based four‐stage system or an anti‐FITC antibody based detection system. Alkaline phosphatase was visualized using a Fast Red naphthol‐capture method and the sections were counterstained with haematoxylin. The results confirm that the method is specific for k or λ mRNA and show that specific mRNAs can be detected in routine formalin‐fixed sections using non‐radioactive techniques with retention of good morphology. The method reliably detects light chain mRNA in cells expressing secretory immunoglobulin. The protocol can also be applied to tissue rich in endogenous biotin by using hapten‐labelled probes. Copyright © 1990 John Wiley & Sons, Ltd.
Authors & Co-Authors
Pringle, J. H.
United Kingdom, Leicester
University of Leicester
Ruprai, Anita K.
United Kingdom, Leicester
University of Leicester
Primrose, Lindsay
United Kingdom, Leicester
University of Leicester
Keyte, John
United Kingdom, Leicester
University of Leicester
Potter, Linda
United Kingdom, Leicester
University of Leicester
Close, Pauline M.
South Africa, Cape Town
University of Cape Town
Lauder, Ian
United Kingdom, Leicester
University of Leicester
Statistics
Citations: 87
Authors: 7
Affiliations: 2
Identifiers
Doi:
10.1002/path.1711620305
ISSN:
00223417
e-ISSN:
10969896
Research Areas
Genetics And Genomics