User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/05/07: Difference between revisions
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====Incubating lysates==== | ====Incubating lysates==== | ||
#incubate 1.5 mL of each sample with 29.3 mg cAMP for 15-30 min. while rotating @ 4 °C | #incubate 1.5 mL of each sample with 29.3 mg cAMP for 15-30 min. while rotating @ 4 °C | ||
#Collect 100 | #Collect 100 μL of these samples | ||
#Put 1.5 mL of each sample on 30 uL of 8-AHA-cAMP beads and incubate 2 – 4 h while rotating @ 4 °C | #Put 1.5 mL of each sample on 30 uL of 8-AHA-cAMP beads and incubate 2 – 4 h while rotating @ 4 °C | ||
=====Washing beads: cAMP precipitation===== | =====Washing beads: cAMP precipitation===== | ||
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==Notes== | ==Notes== | ||
* 50 μL of beads were used instead of 30 μL | * 50 μL of beads were used instead of 30 μL | ||
==Related== | <!-- ==Related== | ||
===Related entries=== | ===Related entries=== | ||
===Same actions=== | ===Same actions=== | ||
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Revision as of 00:32, 21 June 2010
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Summary
Materials & MethodsMaterials
MethodDisrupting cells
Incubating lysates
Washing beads: cAMP precipitation
Samples on gel
Notes
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