In:
Tumori Journal, SAGE Publications, Vol. 83, No. 3 ( 1997-05), p. 673-678
Abstract:
Integrins are widely known cell membrane receptors for extracellular matrix molecules. The β1 integrin subgroup is mainly expressed by kidney cells; immunolocalization of these molecules is usually carried out on cryostatic sections. A commercial monoclonal antibody directed against the human β1 integrin was tested in order to design a method for the detection of this antigen in formalin-fixed, paraffin-embedded human kidney tissue. Methods Specimens obtained from nephrectomies were fixed with 10% formalin and embedded in paraffin. Three different detection protocols were applied after incubation with the anti-human β1 integrin monoclonal antibody (MAB 1977): 1) immunoperoxidase with labeled streptavidin biotin (LSAB), using biotinylated secondary antibodies, peroxidase-labeled biotin-streptavidin, and 3, 3′-diaminobenzidine tetra-hydrochloride (DAB) as the revealing system; 2) immunoperoxidase with tyramide signal amplification (TSA), using biotinylated secondary antibodies, streptavidin-peroxidase, tyramide, streptavidin-peroxidase again and DAB; 3) indirect immunofluorescence with fluorescein-labeled anti-mouse immunoglobulins. Results The best results were obtained with the LSAB detection protocol preceded by a predetection step with proteinase k. Proteinase k pretreatment did not significantly damage the tissue morphology and successfully unmasked 31 integrin antigens. Nonspecific background staining was reduced by a blocking step with swine serum. Similar results were obtained with the TSA detection method; however, although lower concentrations of anti-β1 integrin immunoglobulins and of secondary biotinylated antibody were employed, there was more undesired background staining than with the LSAB protocol. Conclusions The method reported and discussed here may represent a valid tool for research and diagnostic applications based upon detection of β1 integrin in paraffin-embedded human tissues.
Type of Medium:
Online Resource
ISSN:
0300-8916
,
2038-2529
DOI:
10.1177/030089169708300310
Language:
English
Publisher:
SAGE Publications
Publication Date:
1997
detail.hit.zdb_id:
280962-X
detail.hit.zdb_id:
2267832-3
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