Hello,

I am currently trying to visualise microglia trf2 protein in mouse brain sections and cells from culture but not getting any signal for trf2.

I started with blocking in 1xPBS +1% BSA + 0.3% triton X-100 +10% Normal Donkey Serum for 60mins at RT, followed by incubation in fab fragment solution made up of 1x PBS + 0.1% Triton+ 3% serum + 2% BSA + 3% Milk powder and Fab fragment (1:200) for 2hrs. This was followed by incubation in Antibody solution on coverslips O/N at 4 degrees for primary and 60 mins for secondary antibody. This did not give any signal for terf2 but worked quite well for Iba1 microglia marker. I tried modifying this protocol to what I found in the literature and still no positive result and also used 2 different Trf2 antibodies. I have also used as much as 1:10 trf2 concentration and still the same result.

I will so much appreciate if someone can give me a protocol for mouse brain IHC and ICC that worked for them and the specific antibody used.

Thanks