Immunofluorescent Staining of Fixed Mouse Brain Tissue Sections
Katerina Rademacher, Ken Nakamura
Published: 2024-03-30 DOI: 10.17504/protocols.io.kxygx38owg8j/v1
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Abstract
This protocol describes steps for immunofluorescent staining of free floating fixed mouse brain tissue sections.
Steps
1.
Free floating sections in 12-well plates, all steps performed at room temperature.
1.1.
2 x 10min PBS washes.
1.2.
2 x 10min 0.2% PBS-Triton (PBS-T) washes.
1.3.
Block 1hr in blocking buffer.
1.4.
Incubate in primary antibody diluted in blocking buffer overnight.
1.5.
3 x 10min 0.2% PBS-T washes.
1.6.
Incubate in secondary antibody diluted in blocking buffer for 2hrs.
1.7.
2 x 10min PBS-T washes.
1.8.
2 x 10min PBS washes.
1.9.
Mount sections on glass slides, add mounting media and coverslip.
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