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
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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