Background staining of visualization systems in immunohistochemistry: comparison of the Avidin-Biotin Complex system and the EnVision+ system.

Author(s): Vosse BA, Seelentag W, Bachmann A, Bosman FT, Yan P

Publication: Appl Immunohistochem Mol Morphol, 2007, Vol. 15, Page 103-7

PubMed ID: 17536316 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   This study compared the degree of non-specific background staining obtained with two popular detection systems, Envision V+ and the avidin-biotin-complex (ABC) method, for immunohistochemical analysis of formalin-fixed, paraffin-embedded (FFPE) specimens. The following antigens were investigated: Ki-67, cytokeratin, membrane associated antigen, and S-100 protein.

   Summary of Findings:

   ABC and EnVision+ immunohistochemical detection methods were comparable with regard to the intensity and pattern of antigen-specific immunostaining. However, nonspecific staining was not observed with EnVision+ detection while cytoplasmic and nuclear nonspecific background staining was observed with the ABC detection method for multiple biospecimen locations, which was only partially attentuated by an additional step blocking endogenous avidin-binding activity. EnVision+ also produced more robust staining intensity at elevated primary antibody dilutions compared to the ABC method.

   Biospecimens

   • Tissue - Breast
   • Tissue - Kidney
   • Tissue - Larynx
   • Tissue - Liver
   • Tissue - Tonsil
   • Tissue - Ovary
   • Tissue - Pancreas
   • Tissue - Prostate
   • Tissue - Stomach
   • Tissue - Colorectal
   • Tissue - Lung

   Preservative Types

   • Formalin

   Diagnoses:

   • Neoplastic - Carcinoma
   • Normal

   Platform:

   Analyte	Technology Platform
   Protein	Immunohistochemistry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Immunohistochemistry Specific	Detection method	Avidin-biotin complex EnVision+
   Immunohistochemistry Specific	Antibody dilution/concentration	1:50 1:100 1:200 1:300 1:400 1:500
   Immunohistochemistry Specific	Targeted peptide/protein	Ki-67 Cytokeratin Membrane associated antigen S-100 protein
   Biospecimen Acquisition	Biospecimen location	Breast Colon Kidney Larynx Liver Lung Ovary Pancreas Prostate Stomach Tonsil

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