Hello Ainur Botagarova,
We have developed a rapid and efficient proliferating cell nuclear antigen (PCNA) labeling on mouse testicular tissue. Feel free to try / test the protocol on the following article: Article Rapid Screening of Cryopreservation Protocols for Murine Pre...
We used a commercial kit according to the manufacturer's recommendations (Zymed Laboratories, South San Francisco, California). After substrate reaction, stained cells on slides were briefly counterstained with hematoxylin (Merck, Fontenay Sous Bois, France) and mounted with an aqueous mounting medium (Glycergel; Dako, Trappes, France).
It is important to keep in mind that to label proliferating cells it seems to me more appropriate to use Ki67 than PCNA (which also labels cells performing DNA repair).
Hoping that this comment will be useful.
Feel free to discuss or share some informations about your PCNA staining on paraffin fixed slides studies.
Ludovic Dumont, Ph.D.
Dear Ludovic Dumont thank you very much for your useful comment.
I checked the protocol, thank you.
As for the Antibody, I purchased mouse monoclonal antibody (PC10) to PCNA.
I am wondering what kind of the Antigen retrieval you used?
Citrate buffer (pH-6) was used as an antigen retrieval buffer in several papers.
Thank you once again.
Dear Ainur Botagarova,
For PCNA, sections were indeed deparaffinized with xylene, rehydrated in a graded ethanol series, and incubated in 10 mM citrate (pH=6) for 40 minutes at 96°C then left to cool at RT for 20 minutes.
Hoping that this this protocol extension will be useful for your PCNA staining.
Ludovic Dumont, Ph.D.
Dear Ludovic Dumont ,
Thank you for your valuable comments.
With regards,
Ainur Botagarova
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