Formalin-fixed, paraffin-embedded tissues were sectioned at 5-um thickness and stained with hematoxylin and eosin or by IHC as previously described [43 (link)]. Antigen retrieval was achieved by autoclaving in citrate buffer solution (DAKO Target Antigen Retrieval) and IHC for PrPCWD was performed on an automated immunostainer (Ventana Medical Systems) using the monoclonal antibody F99/97.6.1 (VMRD) and Ultraview Red detection kit (Ventana Medical Systems). Positive and negative staining were differentiated based on comparisons with positive and negative deer control tissues included in each run, and negative control tissues showed no immunoreactivity. The extent of PrPCWD deposition in obex sections was graded (- to ++++) using previously published criteria [31 (link)].
Protocol detail
Immunohistochemical Analysis of CWD Prion Deposition
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 : Animal and Plant Quarantine Agency
Other organizations : Canadian Food Inspection Agency
Top 5 similar protocols
Protocol cited in 1 other protocol
Variable analysis
independent variables
- Antigen retrieval method (autoclaving in citrate buffer solution)
- Extent of PrP^CWD deposition (graded - to ++++)
- Tissue samples (formalin-fixed, paraffin-embedded)
- Tissue section thickness (5-um)
- Staining techniques (hematoxylin and eosin, IHC)
- Positive control: Deer tissues with known PrP^CWD positivity
- Negative control: Deer tissues with no PrP^CWD immunoreactivity
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!