The paraffin-embedded tumor sections were stained for anti-Ki67, anti-PCNA, anti-MMP-9, and anti-VEGF (Abcam). Sections (2 µm) were deparaffinized and pretreated with citrate buffer using a heat-induced epitope retrieval protocol. Endogenous peroxidase was blocked with 20% hydrogen peroxide for 15 minutes at room temperature followed by incubation with anti-Ki67, anti-PCNA, anti-MMP-9, and anti-VEGF for 30 minutes, respectively. A biotinylated goat anti-mouse immunglobulin G secondary antibody (Dako, Glostrup, Denmark) was then applied to each slide for 30 minutes. After washing in Tris-hydrochloric acid buffer, the slides were incubated with peroxidase-conjugated streptavidin complex reagent (Dako) and developed with 3,3′-diaminobenzidine for 5 minutes. The slides were counterstained and dehydrated.
Peroxidase conjugated streptavidin complex reagent
Manufactured by Agilent Technologies
Sourced in Denmark
Peroxidase-conjugated streptavidin complex reagent is a laboratory tool that consists of streptavidin, a protein that binds to biotin, coupled with the enzyme peroxidase. This reagent is used in various immunoassay and detection techniques that involve biotin-streptavidin interactions.
Automatically generated - may contain errors
Lab products found in correlation
Anti ki67, by Abcam (3 mentions)
Anti vegf, by Abcam (2 mentions)
Anti caspase 3, by Abcam (2 mentions)
Biotinylated goat anti mouse immunoglobulin g secondary antibody, by Agilent Technologies (2 mentions)
Anti mmp9, by Abcam (1 mentions)
Anti pcna, by Abcam (1 mentions)
Biotin labeled secondary antibody, by Abcam (1 mentions)
Vascular endothelial growth factor (vegf), by Abcam (1 mentions)
4 protocols using peroxidase conjugated streptavidin complex reagent
1
Immunohistochemical Analysis of Tumor Markers
2
Immunohistochemical Analysis of VEGF
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Paraffin sections of the tumor tissue in the nude mice from the control group and the emodin-treated group were obtained, followed by deparaffinization and incubation with 5% goat serum for 2 h. Primary antibody (VEGF; 1:200; Abcam) was then added and incubated at 4°C overnight, followed by incubation with biotin-labeled secondary antibody (Abcam) for 1 h. After washing in Tris-hydrochloric acid butter (TBS), the slides were incubated with peroxidase-conjugated streptavidin complex reagent (Dako, Denmark) and developed with 3,3′-diaminobenzidine for 5 min. The slides were counterstained and dehydrated. Proteins were visualized using microscopy.
3
Immunohistochemical Analysis of Tumor Proliferation
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The paraffin‐embedded tumor sections were stained for anti‐Ki67 and anti‐caspase‐3 (Abcam). Sections (2 μm) were deparaffinized and pretreated with citrate buffer using a heat‐induced epitope retrieval protocol. Endogenous peroxidase was blocked with 20% hydrogen peroxide for 15 minutes at room temperature followed by incubation with anti‐Ki67, and anti‐caspase‐3 for 30 minutes, respectively. A biotinylated goat anti‐mouse immunoglobulin G secondary antibody (Dako, Denmark) was then applied to each slide for 30 minutes. After washing in Tris‐hydrochloric acid buffer (TBS), the slides were incubated with peroxidase‐conjugated streptavidin complex reagent (Dako) and developed with 3,3’‐diaminobenzidine for 5 minutes. The slides were counterstained and dehydrated. Positive cells were detected as a brown staining. The treated specimens were observed and analyzed by using a microscope (OLYMPUS, Japan).
4
Immunohistochemical Analysis of Tumor Markers
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The paraffin‐embedded tumor sections were stained for antiKi67, antiCaspase‐3, and antiVEGF (Abcam, Cambridge, UK). Sections (2 μm) were deparaffinized and pretreated with citrate buffer using a heat‐induced epitope retrieval protocol. Endogenous peroxidase was blocked with 20% hydrogen peroxide for 15 minutes at room temperature followed by incubation with antiKi67, antiCaspase‐3, and antiVEGF for 30 minutes, respectively. A biotinylated goat antimouse immunoglobulin G secondary antibody (Dako, Denmark) was then applied to each slide for 30 minutes. After washing in Tris‐hydrochloric acid buffer (TBS), the slides were incubated with peroxidase‐conjugated streptavidin complex reagent (Dako) and developed with 3,3′‐diaminobenzidine for 5 minutes. The slides were counterstained and dehydrated.
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