Application
Immunohistochemistry free floating
Sample
Rat Tissue sections (brain)
Specification
brain
Other product details
Incubation time
18 hour(s) and 0 minute(s) · Temperature: 4°C
Dilution
1/200
Secondary antibody
Dilution
1/500
Name
Non-Abcam antibody was used: Invitrogen goat anti-mouse AF647
Host species: Mouse
Clonality: Polyclonal
Conjugation: Alexa Fluor® 647
Host species: Mouse
Clonality: Polyclonal
Conjugation: Alexa Fluor® 647
Additional data
Additional Notes
Rats were perfused with 1xPBS and 4% PFA. Organs harvested from IHC-free floating were fixed with 4% PFA for 5-7h (till tissues sink), followed by 15% sucrose overnight and 30% sucrose for 5-7 hours (till tissues sink). All PFA and sucrose fixation process took place in 4 °C fridge. After fixation, rat brains were rapidly freezed with dry ice and sectioned for 30 um/tissue.
30 um rat brain tissues were washed 4x with PBST (3% triton-X) (10 min/wash), then 20 min in H2O2, followed by additional 2x PBST wash. Tissues then were blocked with 2% goat serum for 1h (room temp), then incubated with Anti-NeuN antibody (1:200) for 30min at room temp then 4 °C overnight.
On the following day, well plates with tissues were taken out from 4 °C fridge, incubated with 30 min at room temp, followed by 3x PBST wash. Tissues were then incubated with Invitrogen goat anti-mouse AF647 secondary antibody (1:500) for 3h at room temp, followed by another 3x PBST wash, and were mounted with anti-shade mounting media from vector laboratory.
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提交于 Mar 23 2021