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Procedure for isolating and visualizing hippocampal astrocytes from mice.
Kindly submitted by Gerald Seifert and Christian Steinhäuser, Institute of Cellular Neurosciences, University of Bonn, Germany.
Perform SR101 incubation during ACSF/sucrose incubation at 35°C (Step 3).
ACSF with sucrose
87 mM NaCl
2.5 mM KCl
1.25 mM NaH2PO4
7 mM MgCl2
0.5 mM CaCl2
25 mM NaHCO3
25 mM D-glucose
75 mM sucrose gassed with carbogen
ACSF without sucrose
126 mM NaCl
3 mM KCl
1.25 mM NaH2PO4
2 mM MgSO4
2 mM CaCl2
26 mM NaHCO3
10 mM D-glucose gassed with carbogen
HEPES-buffered solution
150 mM NaCl
5 mM KCl
2 mM MgSO4
2 mM CaCl2
10 mM HEPES
10 mM D-glucose gassed with O2
Adjust solution pH to 7.4 using NaOH or HCl