Glycogen Synthase Assay: Difference between revisions

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[[Category:Glycogen]]

[[Category:Metabolism]]

[[Category:Cell Culture]]

==Materials==

* Glycogen Synthase Buffer ('''GSB''')

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# Incubate 15 min at 37C with occasional shaking

# Place on ice for 15 min.

# Spot 90 uL of reaction mixture onto GF/A filters. ~~Wait 3s and immerse~~ in 70% ethanol ~~at 4C~~

# Spot 90 uL of reaction mixture onto GF/A filters.

# Place in a well of a 6 well dish.

# Add ~2mL of ice cold 70% ethanol.

# Wash 15 min at 4C

# Wash twice 10-20 min at room temperature with fresh 70% ethanol.

# Air dry overnight and count in 5 mL scintillation fluid.

# Air dry overnight on a paper towel and count in 5 mL scintillation fluid. Also count 5 uL of both the - and + G6P solution to calculate total incorporation

@@ Line 1: / Line 1: @@
+[[Category:Glycogen]]
+[[Category:Metabolism]]
+[[Category:Cell Culture]]
 ==Materials==
 * Glycogen Synthase Buffer ('''GSB''')
@@ Line 23: / Line 27: @@
 # Incubate 15 min at 37C with occasional shaking
 # Place on ice for 15 min.
-# Spot 90 uL of reaction mixture onto GF/A filters.  Wait 3s and immerse in 70% ethanol at 4C
+# Spot 90 uL of reaction mixture onto GF/A filters.
+# Place in a well of a 6 well dish.
+# Add ~2mL of ice cold 70% ethanol.
 # Wash 15 min at 4C
 # Wash twice 10-20 min at room temperature with fresh 70% ethanol.
-# Air dry overnight and count in 5 mL scintillation fluid.
+# Air dry overnight on a paper towel and count in 5 mL scintillation fluid.  Also count 5 uL of both the - and + G6P solution to calculate total incorporation