Skip to main content
Journal of Clinical Pathology logoLink to Journal of Clinical Pathology
. 1981 Aug;34(8):859–865. doi: 10.1136/jcp.34.8.859

Immunoperoxidase staining of formalin-fixed, paraffin-embedded, human renal biopsies with a comparison of the peroxidase-antiperoxidase (PAP) and indirect methods.

R A Sinclair, J Burns, M S Dunnill
PMCID: PMC493958  PMID: 7024314

Abstract

The results of immunoperoxidase staining of 33 routinely processed renal biopsies correlated with the results of direct immunofluorescence in 79% of tests performed. Most of the discrepant results were due to positive immunoperoxidase staining, possibly reflecting greater sensitivity of the method. A comparison of two immunoperoxidase methods showed that the indirect method produced less intense staining than the peroxidase-antiperoxidase (PAP) method when equivalent primary antibody titres were used. Lowering the primary antibody titres for the indirect technique resulted in equivalent staining by the two methods. Predigestion by protease VII effectively revealed the antigens under study without causing tissue damage or section loss.

Full text

PDF
859

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bergroth V., Reitamo S., Konttinen T., Lalla M. Sensitivity and nonspeicific staining of various immunoperoxidase techniques. Histochemistry. 1980;68(1):17–22. doi: 10.1007/BF00498496. [DOI] [PubMed] [Google Scholar]
  2. Burns J. Background staining and sensitivity of the unlabelled antibody-enzyme (PAP) method. Comparison with the peroxidase labelled antibody sandwich method using formalin fixed paraffin embedded material. Histochemistry. 1975 Jun 5;43(3):291–294. doi: 10.1007/BF00499711. [DOI] [PubMed] [Google Scholar]
  3. Clausen P. P. Immunohistochemical demonstration of alpha-1-antitrypsin in liver tissue. A methodological investigation. Acta Pathol Microbiol Scand A. 1980 Sep;88(5):299–306. doi: 10.1111/j.1699-0463.1980.tb02500.x. [DOI] [PubMed] [Google Scholar]
  4. Davey F. R., Busch G. J. Immunohistochemistry of glomerulonephritis using horseradish peroxidase and fluorescein-labeled antibody: a comparison of two technics. Am J Clin Pathol. 1970 Apr;53(4):531–536. doi: 10.1093/ajcp/53.4.531. [DOI] [PubMed] [Google Scholar]
  5. Elias J. M., Miller F. A comparison of the unlabeled enzyme method with immunofluorescence for the evaluation of human immunologic renal disease. Am J Clin Pathol. 1975 Oct;64(4):464–471. doi: 10.1093/ajcp/64.4.464. [DOI] [PubMed] [Google Scholar]
  6. Heyderman E. Immunoperoxidase technique in histopathology: applications, methods, and controls. J Clin Pathol. 1979 Oct;32(10):971–978. doi: 10.1136/jcp.32.10.971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. MacIver A. G., Giddings J., Mepham B. L. Demonstration of extracellular immunoproteins in formalin-fixed renal biopsy specimens. Kidney Int. 1979 Nov;16(5):632–636. doi: 10.1038/ki.1979.174. [DOI] [PubMed] [Google Scholar]
  8. Mason D. Y., Biberfeld P. Technical aspects of lymphoma immunohistology. J Histochem Cytochem. 1980 Aug;28(8):731–745. doi: 10.1177/28.8.6160179. [DOI] [PubMed] [Google Scholar]
  9. Mepham B. L., Frater W., Mitchell B. S. The use of proteolytic enzymes to improve immunoglobulin staining by the PAP technique. Histochem J. 1979 May;11(3):345–357. doi: 10.1007/BF01005033. [DOI] [PubMed] [Google Scholar]
  10. Sternberger L. A., Hardy P. H., Jr, Cuculis J. J., Meyer H. G. The unlabeled antibody enzyme method of immunohistochemistry: preparation and properties of soluble antigen-antibody complex (horseradish peroxidase-antihorseradish peroxidase) and its use in identification of spirochetes. J Histochem Cytochem. 1970 May;18(5):315–333. doi: 10.1177/18.5.315. [DOI] [PubMed] [Google Scholar]
  11. Tubbs R. R., Gephardt G., Valenzuela R., Deodhar S. An approach to immunomicroscopy of renal disease with immunoperoxidase and periodic acid-Schiff counterstain (IMPAS stain). Am J Clin Pathol. 1980 Feb;73(2):240–244. doi: 10.1093/ajcp/73.2.240. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Pathology are provided here courtesy of BMJ Publishing Group

RESOURCES