Testis squashes and immunostaining were performed as previously described (Kearse et al., 2011 (link)). Briefly, testis tissue was squashed with a #1 coverslip onto a glass slide and immediately frozen on dry ice. Tissue was fixed in ice-cold ethanol and 4% formaldehyde and washed two times in PBST with sodium deoxycholate (1X PBS, 0.3% sodium deoxycholate, 0.3% Triton X-100) and one time in PBST. Tissue was then incubated overnight in primary antibody, washed four times in PBTB (1X PBS, 0.1% bovine serum albumin [BSA], 0.1% Triton X-100), and incubated for 1 h in secondary antibody. Tissue was washed four times in PBTB, followed by two times in 1X PBS and mounted using Fluormount-G (Southern Biotech; #0100-01). Imaging was completed using a Nikon Eclipse TE200U.
Protocol detail
Testis Immunostaining Protocol
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.
Antibody
Bovine serum albumin
Cold
Dry ice
Ethanol
Formaldehyde
Frozen
Sodium deoxycholate
Squashes
Testis
Tissue
Triton x 100
Corresponding Organization :
Other organizations : Lehigh University
Top 5 similar protocols
Variable analysis
independent variables
- Testis tissue squashing
- Incubation in primary antibody
- Incubation in secondary antibody
- Imaging of testis tissue
- Tissue fixation in ice-cold ethanol and 4% formaldehyde
- Tissue washing in PBST and PBTB
- Mounting using Fluormount-G
- Imaging using Nikon Eclipse TE200U
- No positive or negative controls were explicitly mentioned in the provided information.
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!