Sections of formalin-fixed, paraffin-embedded tissues (4 μm thick) were de-paraffinized, subjected to heat-mediated antigen retrieval in EDTA buffer (pH 9.0) at 98 °C for 40 min, and then blocked in 2.5% normal horse serum (ImmPRESS Horse Anti-Rabbit IgG Polymer kit; Vector Laboratories, Riverside, CA, USA) at room temperature for 20 min. They were incubated with anti-CD163 antibody (Abcam) overnight at 4 °C, followed by incubation with the appropriate secondary antibody (ImmPRESS Horse Anti-Rabbit IgG Polymer kit; Vector Laboratories) at room temperature for 30 min, and then incubated with 3,3′-diaminobenzidine (Sigma-Aldrich) at room temperature for 5 min. After that, they were heated again in EDTA buffer in the same way. They were then blocked, followed by incubation with anti-CD68 antibody (Abcam) and the secondary antibody in the same way, and next incubated with working solution prepared with Vector SG Peroxidase (HRP) Substrate Kit (Vector Laboratories) at room temperature for 5 min.
Vector sg peroxidase hrp substrate kit
Manufactured by Vector Laboratories
Sourced in United States
The Vector SG Peroxidase (HRP) Substrate Kit is a laboratory product that provides a substrate for the detection of horseradish peroxidase (HRP) enzyme in various immunochemical and biochemical applications. The kit contains a ready-to-use solution that produces a blue-green colored product upon reaction with the HRP enzyme.
Automatically generated - may contain errors
Lab products found in correlation
Immpress horse anti rabbit igg polymer kit, by Vector Laboratories (2 mentions)
3 3 diaminobenzidine (dab), by Merck Group (2 mentions)
Anti cd163 antibody, by Abcam (1 mentions)
Anti cd68 antibody, by Abcam (1 mentions)
Ab28364, by Abcam (1 mentions)
Vector nuclear fast red, by Vector Laboratories (1 mentions)
Anti pro spc, by Merck Group (1 mentions)
Anti foxp3 antibody, by Abcam (1 mentions)
Anti cd3 antibody, by Abcam (1 mentions)
4 protocols using vector sg peroxidase hrp substrate kit
1
Immunohistochemical Analysis of CD163 and CD68
2
Murine Vascular Endothelial Cell Immunostaining
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Murine vascular blood vessels were detected using species-specific CD31 immunostaining. Unstained tumor sections were processed after conventional deparaffinization and endogenous peroxidase quenching using 3% hydrogen peroxide. Tumor tissue sections were incubated with 10% normal donkey serum in PBS for 3 h at room temperature to block non-specific binding sites. Rabbit anti-mouse CD31 primary antibody (ab28364, Abcam, UK) was diluted 1: 50 in 2% donkey serum in PBS and incubated for 3 h at room temperature, followed by incubation with the secondary anti-rabbit HRP using an X-Cell-Plus HRP Detection Kit (Menarini Diagnostics, UK) for 45 min at room temperature. The reaction was revealed using a VECTOR SG Peroxidase (HRP) Substrate Kit (Vector Laboratories, US) for 10 min at room temperature. All sections were counterstained with Neutral Fast Red (Sigma Aldrich, UK).
3
Immunohistochemical Staining of Kir7.1 and pro-SPC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sections (4 μm) were treated with 10% hydrogen peroxide, blocked with 2.5% normal horse serum, and incubated with anti-Kir7.1 [[5 (link)], 1:15,000] or anti-pro-SPC (1: 2,000; Millipore) overnight at 4°C. Detection of antigen/antibody complexes was performed with ImmPRESSTM (Peroxidase) Polymer Anti-Rabbit Ig Reagent (Vector Laboratories, USA). The reaction was visualized using Vector SG Peroxidase (HRP) Substrate Kit (Vector Laboratories, USA). In controls the incubation with primary antibody was omitted. Sections were counterstained with Vector Nuclear Fast Red (Vector Laboratories, USA).
4
Dual Immunohistochemical Staining Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Double immunostaining was performed according to the following methods. Sections of formalin-fixed, paraffin-embedded tissues (4 μm thick) were de-paraffinized, subjected to heat-mediated antigen retrieval in citric acid buffer at 98 °C for 40 min, and then blocked in 5% skim milk at room temperature for 1 h. They were incubated with anti-FOXP3 antibody (Abcam) overnight at 4 °C, followed by incubation with the appropriate secondary antibody (DAKO) at room temperature for 1 h, and then incubated with 3,3′-diaminobenzidine (Sigma-Aldrich) at room temperature for 5 min. After that, they were heated again in EDTA buffer (pH 9.0) in the same way. They were then blocked in 2.5% normal horse serum (ImmPRESS Horse Anti-Rabbit IgG Polymer kit; Vector Laboratories, Riverside, CA, USA) at room temperature for 20 min, followed by incubation with anti-CD3 antibody (Abcam) overnight at 4 °C. They were incubated with the secondary antibody (ImmPRESS Horse Anti-Rabbit IgG Polymer kit; Vector Laboratories) at room temperature for 30 min and then incubated with working solution prepared with Vector SG Peroxidase (HRP) Substrate Kit (Vector Laboratories) at room temperature for 5 min.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!