Synaptosomes on coverslips were fixed (4% Paraformaldehyde, 4% sucrose, 1× PBS) for 10 min at room temperature, washed three times with PBS for 5 min and then stored at 4 °C until use. Synaptosomes were blocked and permeabilized with PGT buffer (PBS, 2 g/L gelatin, 0.25% Triton X-100 and when needed 5% normal goat serum) and subsequently incubated with primary antibodies in PGT buffer (1 h at room temperature), washed three times with PGT and incubated with secondary antibodies in PGT (1 h at room temperature). Three final washes with PGT buffer were performed prior to a washing step in 1× PBS and a final rinse in ultrapure water. Coverslips were mounted on glass slides with Fluoromount-G mounting solution (Sigma) and stored at 4 °C until observation.
Fluoromount g mounting solution
Manufactured by Merck Group
Fluoromount-G is a mounting solution designed for use in microscopy applications. It is a water-based, non-fluorescent medium that can be used to mount and preserve fluorescent-labeled specimens for analysis.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using fluoromount g mounting solution
1
Immunocytochemistry of Synaptosomes
2
Synaptosome Immunofluorescence Staining
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Synaptosomes on coverslips were fixed (4% paraformaldehyde, 4% sucrose, PBS) for 10min at room temperature, washed three times with PBS for 5min and then stored at 4°C until use. Synapstosomes were blocked and permeabilized with PGT buffer (PBS, 2g/L gelatin, 0.25% Triton X-100) and subsequently incubated with primary antibodies in PGT buffer (1h at room temperature), washed 3 times with PGT and incubated with secondary antibodies in PGT (1 hour at room temperature). Three final washes with PGT buffer were performed prior to a washing step in PBS and a final rinse in ultrapure water. Coverslips were mounted on glass slides with Fluoromount-G mounting solution (Sigma) and stored at 4°C until observation.
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