Immunohistochemical staining was performed on paraffin-embedded mouse tissue sections (5 mm) to determine Ki-67 and Notch1 expression. The slides were incubated with anti–Ki-67 and anti-Notch1 antibodies (1:500, Abcam) overnight at 4°C. A horseradish peroxidase (HRP) detection system (ZSGB-Bio, Beijing, China) and the diaminobenzidine (DAB) substrate kit (ZSGB-Bio) were used as detection reagents. After counterstaining with hematoxylin (ZSGB-Bio), the sections were dehydrated and mounted, and observed under a light microscope (Olympus, Tokyo, Japan). The positive rates were measured using Image-Pro Plus v.6.0 software (Media Cybernetics, Bethesda, MD, USA).
Diaminobenzidine dab substrate kit
Manufactured by ZSGB-BIO
Sourced in China
Diaminobenzidine (DAB) substrate kit is a laboratory reagent used for the detection and visualization of target proteins in immunohistochemistry and immunocytochemistry applications. It provides a brown-colored precipitate at the site of the antigen-antibody reaction, allowing for the identification and localization of the protein of interest.
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3 protocols using diaminobenzidine dab substrate kit
1
Immunohistochemical Analysis of Ki-67 and Notch1
2
Quantifying Ki-67 Expression in Mouse Tissues
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Immunohistochemical staining was performed on paraffin-embedded mouse tissue sections (5 mm) to determine Ki-67 expression. The slides were incubated with anti-Ki-67 antibody (1:500, Abcam (ab15580)) overnight at 4°C. Horseradish peroxidase (HRP) Detection System (ZSGB-bio, Beijing, China) and diaminobenzidine (DAB) Substrate Kit (ZSGB- bio) were used as detection reagents. After counterstaining with hematoxylin (ZSGB-bio), the sections were dehydrated and mounted, and observed under light microscopy (Olympus, Tokyo, Japan). The positive rates were measured using Image-Pro Plus v. 6.0 software (Media Cybernetics, Bethesda, MD, USA). All reactions were performed in triplicate.
3
Endometriosis Immunohistochemistry Protocol
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Paraffin sections of ectopic and eutopic endometrial tissue from patients with ovarian-type endometriosis and normal endometrial tissue from patients without endometriosis were collected. The sections were incubated at 65 °C for 2 h, deparaffinised with xylene, and hydrated with ethanol. Antigen repair was performed using high-pressure boiling, followed by blocking of endogenous peroxidase blocker (ZSGB, Beijing, China) and incubation with anti-IGFBP1 primary antibody (Proteintech, Wuhan, China) (1:200) at 4 °C overnight. Next, sections were incubated with HRP-labelled secondary antibody (ZSGB, Beijing, China) for 30 min. Colour was developed using the diaminobenzidine (DAB) substrate kit (ZSGB, Beijing, China) for 1 to 2 min, and nuclei were re-stained with haematoxylin.
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