I'm also trying to stain for these markers, but on mice skin sections. I have also trouble with p16-staining, but it can have to do something with the antibodies... In articles you'll see that they stained with p16-antibodies from santa cruz, but they are not being sold anymore. The antibodies from other companies for p16 are unfortunately not useful for staining. Which antibodies (company and clones) are you using?

Hi Marloes Jonkhout !

We used p16 and p21 antibodies from Santa Cruz for mice sections, but without any success.

Hi Carolina,

It is a common problem when you are dealing with paraffin-embedded sections and brain is more sensitive to processing steps and antigen masking.

Try to us different PH of your antigen retrieval solution. For example try PH 9, 7, 6 and 4.

Try also to change the heating mechanism, if you microwave, change to autoclave at 121 c for 1 min.

Of course try different antibody concentrations.

This should work hopefully

Hi Elsayed Mohammed ,

For the antigen retrieval, the best pH until now is 9, but I only tried 9 and 6.

We usually use a water bath as a heating mechanism at 97ºC for 20min, we never use the autoclave, do you think that will make a significant difference?

Thank you for your suggestions!