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==Materials==
* '''Homogenization Buffer''' (50 mM Tris, 5 mM EDTA, 30 mM Mannitol, PI inhibitor)
* 10M KOH
* '''Chloroform/Methanol Mixture''' (2:1)
* '''Butanol Mixture''': 3 mL butanol, 1.66 mL Triton-X114, 0.33 mL Methanol
* Sigma Triglyceride Assay Kit (Cat 337-B)
==Protocol==
# Weigh out 200-500mg tissue (record weight for normalization)
# Homogenize with dounce homogenizer in 2 mL '''Homogenization Buffer'''.
# Remove 200 uL to a tube containing 5 uL KOH
# Mix by inverting
# Add 800 uL '''Chloroform/Methanol Mixture'''
# Vortex vigorously then sit at room temperature for 5 min
# Centrifuge 10min at 13 000 RPM
# Take 180 uL of the bottom layer into a fresh tube.
# Dry in fume hood overnight (or until completely dry)
# Add 50uL of '''Butanol Mixture'''
# Measure triglyceride levels using Sigma Diagnostic Kit using 5 uL of sample.
* '''Homogenization Buffer''' (50 mM Tris, 5 mM EDTA, 30 mM Mannitol, PI inhibitor)
* 10M KOH
* '''Chloroform/Methanol Mixture''' (2:1)
* '''Butanol Mixture''': 3 mL butanol, 1.66 mL Triton-X114, 0.33 mL Methanol
* Sigma Triglyceride Assay Kit (Cat 337-B)
==Protocol==
# Weigh out 200-500mg tissue (record weight for normalization)
# Homogenize with dounce homogenizer in 2 mL '''Homogenization Buffer'''.
# Remove 200 uL to a tube containing 5 uL KOH
# Mix by inverting
# Add 800 uL '''Chloroform/Methanol Mixture'''
# Vortex vigorously then sit at room temperature for 5 min
# Centrifuge 10min at 13 000 RPM
# Take 180 uL of the bottom layer into a fresh tube.
# Dry in fume hood overnight (or until completely dry)
# Add 50uL of '''Butanol Mixture'''
# Measure triglyceride levels using Sigma Diagnostic Kit using 5 uL of sample.