Samples were stained with H&E, ERG, and PIN-4, and laser capture microdissected (LCM) as previously described (13 (link)). No differences were observed between H&E, ERG, and PIN-4 staining for tissues fixed in formalin versus PAXgene. For c-Myc and MAD automated immunohistochemical staining, 5 μm FFPE sections were baked for 1 hour at 60°C. After cooling to room temperature, antigen retrieval was performed using PT Link (Dako). Staining was with EnVision FLEX High pH kit (Dako, cat# K8000021-2) following the manufacturer's recommendations. Sections were incubated with 300 μL primary antibody (anti-c-Myc, Abcam cat# ab32072, 1:200 dilution; anti-MAD, Sigma-Aldrich cat# HPA001599, 1:1000 dilution) for 60 minutes at room temperature.
Protocol detail
Immunohistochemical Staining Protocol for Tissue Analysis
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.
Corresponding Organization : Harvard University
Other organizations : National Cancer Institute, Center for Cancer Research, Emory University, Children's Cancer Center, Baylor College of Medicine
Top 5 similar protocols
Variable analysis
independent variables
- Tissue fixation method (formalin vs. PAXgene)
- Staining characteristics of H&E, ERG, and PIN-4
- Sections of 5 μm FFPE for c-Myc and MAD staining
- Baking of sections for 1 hour at 60°C
- Antigen retrieval using PT Link (Dako)
- Staining with EnVision FLEX High pH kit (Dako)
- Incubation with primary antibodies (anti-c-Myc, anti-MAD) for 60 minutes at room temperature
- Positive control: Not specified
- Negative control: Not specified
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!