Staining was performed as previously reported (39 (link)). In brief, tissue sections were cut at 4μm and deparaffinized in xylene and absolute alcohol. Endogenous peroxidase activities were blocked with a solution of 3% hydrogen peroxide and absolute alcohol 1:1 for 15 minutes. Heat induced antigen retrieval was performed with a pressure cooker (122+/−2°C) for 45 seconds at 15+/−5 PSI in citrate buffer (pH6.0). Immunohistochemistry was performed using a polyclonal rabbit antibody to FOXJ1 (HPA005714, Sigma-Aldrich) at a dilution of 1:75 incubated for 45 minutes at room temperature followed by labeled polymer-HRP anti-rabbit IgG incubated 30 minutes and visualized with 3,3′-Diaminobenzidine (DAB) (brown in color: Dako Envision System). The slides were counterstained with hematoxylin. Images were acquired using an Olympus BX41 microscope and an Olympus DP26 digital camera.
Protocol detail
Immunohistochemical Staining of FOXJ1 in Tissue Sections
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Absolute alcohol
Anti igg
Antibody
Antigen
Buffer
Camera
Citrate
Dilution
Hematoxylin
Hydrogen 3
Immunohistochemistry
Microscope
Peroxidase
Peroxide
Polymer
Pressure
Rabbit
Seconds
Tissue
Xylene
Corresponding Organization :
Other organizations : Brigham and Women's Hospital, Harvard University, Boston Children's Hospital, Johns Hopkins University, Dana-Farber Cancer Institute
Top 5 similar protocols
Protocol cited in 1 other protocol
Variable analysis
independent variables
- Staining protocol (as previously reported)
- Expression of FOXJ1 protein
- Tissue sections cut at 4μm
- Deparaffinization in xylene and absolute alcohol
- Blocking of endogenous peroxidase activities with 3% hydrogen peroxide and absolute alcohol 1:1 for 15 minutes
- Heat-induced antigen retrieval in a pressure cooker (122+/−2°C) for 45 seconds at 15+/−5 PSI in citrate buffer (pH6.0)
- Incubation with polyclonal rabbit antibody to FOXJ1 (HPA005714, Sigma-Aldrich) at a dilution of 1:75 for 45 minutes at room temperature
- Incubation with labeled polymer-HRP anti-rabbit IgG for 30 minutes
- Visualization with 3,3′-Diaminobenzidine (DAB)
- Counterstaining with hematoxylin
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
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!