Kidney tissues were fixed with paraformaldehyde, embedded in paraffin, and cut into sections for immunostaining with antibody against PPARγ. Subsequently, sections were deparaffinized using a graded series of ethanol, and stained using primary antibody against PPARγ. After washing with PBS, sections were incubated with EnVision+/HRP/Rb (DAKO, Glostrup, Denmark) for 30 min at room temperature. The staining was visualized using 3, 3’-diaminobenzidine (DAB) substrate and then counterstained with hematoxylin for 30 s. All sections were photographed using an Olympus BH2 microscope (Olympus).
Envision hrp rb
Manufactured by Agilent Technologies
Sourced in Denmark
The EnVision+/HRP/Rb is a lab equipment product from Agilent Technologies. It is a detection system that utilizes horseradish peroxidase (HRP) and rabbit (Rb) antibodies for sensitive signal amplification in various assays.
Automatically generated - may contain errors
5 protocols using envision hrp rb
1
Immunohistochemical Analysis of PPARγ Expression in Kidney Tissues
2
Immunohistochemical Analysis of PPAR-α in Lung Tissue
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Lungs were fixed in 4% paraformaldehyde, embedded in paraffin and cut into sections of 4 μm in thickness. Sections were incubated with primary antibodies against PPAR-α (Cell Signaling Technology). After washing with phosphate-buffered saline (PBS), these sections were incubated with EnVision+HRP/Rb (DAKO, Glostrup, Denmark) at room temperature for 30 minutes. The positive signals of tissue sections were detected using the DAB (3,3′-diaminobenzidine) substrate kit (ZSGB-BIO, China) following the manufacturer’s instructions. The images were caught under a light microscope (Nikon, Tokyo, Japan).
3
Immunohistochemical Analysis of β-Catenin
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Segments were fixed with 4% paraformaldehyde overnight at 4 °C, embedded in paraffin and cut at a thickness of 4 μm. Sections were incubated with primary antibodies against β-catenin (Cell Signaling Technology). After washing with PBS, tissue sections were incubated with EnVision+/HRP/Rb (DAKO, Glostrup, Denmark) for 30 min at room temperature. The sections were then incubated in 3, 3'-DAB (Maxim, Fuzhou, China) for 5 min and then counterstained with hematoxylin for 30 s. All sections were photographed with a Nikon TE2000-U camera (Nikon, Tokyo, Japan) equipped with Nikon optics.
4
Lysozyme Immunohistochemistry Protocol
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Epitopes were retrieved using boiled 0.01 M citrate buffer (pH 6.0). All sections were incubated with anti-mouse Lyz antibody (1: 2,500; Abcam, USA) diluted with PBS at 4°C overnight, and then incubated with EnVision+/HRP/Rb (DAKO, Denmark) for 30 min. The sections were incubated in 3, 3′-diaminobenzidine tetrahydrochloride (Maxin, China) for 5 min and then counterstained with hematoxylin for 30 s.
5
Immunohistochemical Analysis of Tumor Tissue
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Tumor segments were fixed with 4% paraformaldehyde overnight at 4°C and subsequently embedded in paraffin. The segments were then sectioned to a thickness of 4 μm. Dewaxing procedures were followed by blocking non‐specific sites with a 5% BSA solution for 1 h. Subsequently, the sections were incubated overnight at 4°C with primary antibodies against Ki‐67 (Wanleibio, Shenyang, China) or PLIN3 (Proteintech, Wuhan, China) and washed with PBS. EnVision+/HRP/Rb (DAKO, Glostrup, Denmark) was applied for 1 h at 37°C. The segments were then covered with 3,3′‐Diaminobenzidine (3,3′‐DAB, Maxim, Fuzhou, China) for 5 min and counterstained with hematoxylin for 30 s. Finally, sections were photographed with a TE2000‐U camera (Nikon, Tokyo, Japan).
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