Mar 29, 2024

Public workspaceImmunofluorescent Staining of Fixed Mouse Brain Tissue Sections

DOI
dx.doi.org/10.17504/protocols.io.kxygx38owg8j/v1
  • Katerina Rademacher1,
  • Ken Nakamura1
  • 1Gladstone Institute of Neurological Disease
Haru Yamamoto
Open access
DOI: dx.doi.org/10.17504/protocols.io.kxygx38owg8j/v1
Protocol Citation: Katerina Rademacher, Ken Nakamura 2024. Immunofluorescent Staining of Fixed Mouse Brain Tissue Sections. protocols.io https://dx.doi.org/10.17504/protocols.io.kxygx38owg8j/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: January 07, 2024
Last Modified: May 31, 2024
Protocol Integer ID: 93032
Keywords: ASAPCRN
Funders Acknowledgement:
ASAP
Grant ID: 020529
Abstract
This protocol describes steps for immunofluorescent staining of free floating fixed mouse brain tissue sections.
Materials
  • 1X PBS (UCSF Stem Cell Core)
  • Triton X-100 (Sigma Aldrich 93443)
  • Fetal bovine serum (UCSF Stem Cell Core)
  • Primary antibody
  • Secondary antibody
  • Blocking buffer: 0.2% Triton, 4% FBS in 1X PBS
  • Glass microscope slides (Fisher 22-038-103)
  • Anti-fade mounting medium with or without DAPI (Vector Laboratories H140010)
Free floating sections in 12-well plates, all steps performed at room temperature.
2 x 10min PBS washes.
2 x 10min 0.2% PBS-Triton (PBS-T) washes.
Block 1hr in blocking buffer.
Incubate in primary antibody diluted in blocking buffer overnight.
3 x 10min 0.2% PBS-T washes.
Incubate in secondary antibody diluted in blocking buffer for 2hrs.
2 x 10min PBS-T washes.
2 x 10min PBS washes.
Mount sections on glass slides, add mounting media and coverslip.