Immunohistochemistry was performed on 5 microns thick sections of formalin-fixed and paraffin-embedded tissue samples. Sections were picked on charged glass slides and deparaffinised, hydrated, then treated for antigen retrieval at a high pH (pH 8) using an automated immunostainer (Dako, Denmark). Rabbit polyclonal anti-CD117 (c-kit) antibodies (CD 117, Cat. No. A4502, Dako, Denmark) at dilution 1:200 was used to label the interstitial cells. Goat anti-rabbit biotinylated immunoglobulins/HRP (Cat. No. P0448, Dako, Denmark) were used at dilution 1:300. Streptavidin–biotin–peroxidase complex and peroxidase-DAB (3,3’diaminobenzidine) (Dako, Denmark) detection method was preformed according to the manufacturer’s instructions. Sections were counterstained with Mayer’s haematoxylin. Positive and negative control slides were included in each run. As a negative control, a tissue section was processed as described but with the primary antibody omitted. Minimal of 10 fields were examined in each section at different magnification, and a semiquantitative analysis of distribution of ICCs. The pathologist was blended for the sample origin being BOO or sham-normal and 6 or 8 weeks duration of BOO.
Streptavidin biotin peroxidase complex and peroxidase dab 3 3 diaminobenzidine
Manufactured by Agilent Technologies
Sourced in Denmark
The Streptavidin–biotin–peroxidase complex is a reagent that utilizes the high-affinity interaction between streptavidin and biotin to form a complex with horseradish peroxidase (HRP). This complex can be used as a detection system in various bioanalytical techniques. The peroxidase-DAB (3,3'diaminobenzidine) system is a chromogenic substrate for peroxidase enzymes, which can be used to visualize and quantify the presence of target analytes in a sample.
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2 protocols using streptavidin biotin peroxidase complex and peroxidase dab 3 3 diaminobenzidine
1
Immunohistochemical Analysis of ICC in BOO
2
Immunohistochemical Labeling of Cajal Cells
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Immunohistochemistry was performed on 5 microns thick sections of formalin-fixed and paraffin-embedded tissue samples. Sections were picked on charged glass slides and deparaffinised, hydrated, then treated for antigen retrieval at a high pH (pH 8) using an automated immunostainer (Dako, Denmark). Rabbit polyclonal anti-CD117 (c-kit) antibodies (CD 117, Cat. No. A4502, Dako, Denmark) at dilution 1:200 were used to label the Cajal cells. Goat anti-rabbit biotinylated immunoglobulins/HRP (Cat. No. P0448, Dako, Denmark) were used at dilution 1:300. Streptavidin-biotin-peroxidase complex and peroxidase-DAB (3,3'diaminobenzidine) (Dako, Denmark) detection method was preformed according to the manufacturer's instructions. Sections were counterstained with Mayer's haematoxylin. Positive and negative control slides were included in each run. As a negative control, a tissue section was processed as described but with the primary antibody omitted.
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