Related topics: Alphabetical list, Neurodegeneration, Neurotransmission
On August 15, 2019
Trine Rasmussen wrote:
This antibody is very good for fluorescence IHC staining of α synuclein on free floating brain sections.
Mice were injected with α-synuclein AAV vector in the right hemisphere (striatum). The left hemisphere was left untreated. After 4 months the mice were sacrificed. Their brains were removed and fixed in PFA and cut into 20 µm sections. The tissue was stained using a free floating immunohistochemistry protocol. Clear streaks of stained α-synuclein were visible at the site of injection compared to control.
Sections were collected in a multi-well plate with 300 µl 1x target retrieval solution (Dako, #S1699), incubated at 70 °C for 30 minutes and allowed cool at room temperature.
The sections were rinsed for 2 minutes in water, 3×10 minutes in TBS and 30 minutes in 1%BSA/0.3% Triton X/TBS and allowed to incubate with the 1:500 α synuclein antibody in TB buffer (50 mM) overnight at 4 °C with gentle agitation.
The staining was left at room temperature for 1 hour and subsequently rinsed 3×10 minutes in TBS. Sections were incubated with 1:300 secondary antibody in TB (50 nM) for 4 hours at room temperature. The sections were rinsed 3×10 minutes in TBS with hoechst added the last 10 minutes.
Sections were mounted on slides and allowed to dry.
Pictures were obtained using confocal microscopy.