Jul 12, 2024
Immunostaining- Fluorescent
- daniel.dautan daniel1,2,
- Per Svenningsson1,2
- 1Department of Clinical Neuroscience, Karolinska Institutet, 171 76 Stockholm, Sweden;
- 2Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, MD 20815, USA
Protocol Citation: daniel.dautan daniel, Per Svenningsson 2024. Immunostaining- Fluorescent. protocols.io https://dx.doi.org/10.17504/protocols.io.j8nlk88y6l5r/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: June 04, 2024
Last Modified: July 12, 2024
Protocol Integer ID: 101197
Keywords: ASAPCRN, immunofluorescence, mouse, brain,
Funders Acknowledgement:
Aligning Science Across Parkinson's
Grant ID: 020608
Abstract
Staining of mouse brain sections with immunofluroescent visualization.
Wash freshly sectioned slices of tissue 2-3 times with 1X PBS to remove OCT.
Block with 5% Normal donkey/Goat serum in 0.3% TRITON X-100 in 1X PBS for 01:00:00 at Room temperature .
1h
Wash sections 5 times in 1X PBS.
Following washes, transfer sections to a primary antibody solution containing a mix of primary antibody diluted in 0.3% TRITON X-100 in 1X PBS with 1% Normal Donkey/Goat serum Overnight at 4 °C .
1h
The following day, wash sections 5 times in 1X PBS.
Transfer sections to secondary solution containing the adequate secondary antibody diluted in 0.3%
TRITON X-100 in 1X PBS with 1% Normal Donkey/Goat serum for 04:00:00 atRoom temperature .
4h
Wash sections 3 times in 1X PBS.
Mount sections on microscope slides with medium (Vectashield with 4',6-diamidino-2-phenylindole (DAPI), H-1800-10).