Immunohistochemical staining was performed using a two-step method with heat-induced antigen-retrieval procedures (14 (link)). The cancer tissues were fixed in 4% paraformaldehyde. Following dehydration, the tissue was sliced and blocked in 10% BSA (Sigma-Aldrich; Merck KGaA). The following primary antibodies were used: Rabbit anti-human CCBE1 monoclonal antibody (cat. no. HPA041361, 1:50-1:200; Sigma-Aldrich, Merck KGaA) and rabbit anti-human LYVE1 monoclonal antibody (cat. no. ab33682, 1:100; Abcam). The slices (20 µm) were incubated with horseradish peroxidase-conjugated secondary antibodies (1:100, peroxidase conjugated goat anti-rabbit IgG, TA130023; OriGene Technologies, Inc.) at room temperature for 2 h. The primary antibody was applied in the negative control. The images were visualized by light microscope (Olympus Corporation, Tokyo, Japan).
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Immunohistochemical Staining of CCBE1 and LYVE1 in Cancer Tissues
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Anti antibody
Anti igg
Antibodies
Antibody
Antigen
Cancer
Dehydration
Goat
Horseradish peroxidase
Human
Microscope light
Paraformaldehyde
Peroxidase
Rabbit
Tissue
Corresponding Organization :
Other organizations : Southern Medical University, Nanjing University, Nanjing General Hospital of Nanjing Military Command, Jiangsu University
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Variable analysis
independent variables
- Heat-induced antigen-retrieval procedures
- Immunohistochemical staining
- Fixed in 4% paraformaldehyde
- Dehydration
- Sliced
- Blocked in 10% BSA (Sigma-Aldrich; Merck KGaA)
- Primary antibodies: Rabbit anti-human CCBE1 monoclonal antibody (cat. no. HPA041361, 1:50-1:200; Sigma-Aldrich, Merck KGaA) and rabbit anti-human LYVE1 monoclonal antibody (cat. no. ab33682, 1:100; Abcam)
- Horseradish peroxidase-conjugated secondary antibodies (1:100, peroxidase conjugated goat anti-rabbit IgG, TA130023; OriGene Technologies, Inc.)
- Incubated at room temperature for 2 h
- Primary antibody applied in the negative control
- Visualized by light microscope (Olympus Corporation, Tokyo, Japan)
- primary antibody applied in the negative control
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