Triglyceride Assay from Cells and Tissues: Difference between revisions

Mpeloqu1 (talk | contribs)
Updated protocol for tissue culture
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==Protocol==
==Protocol==
#Weigh out 30mg tissue (record weights for normalization) on dry ice into round bottom eppendorf tube. Add one stainless steel ball bearing.
#Weigh out 30mg tissue (record weights for normalization) on dry ice into round bottom eppendorf tube. Add one stainless steel ball bearing.
#Add 500ul Homogenization Buffer
#Add 200ul Homogenization Buffer
#Homogenize with Qiagen Tissue Lyser for 3 minutes @ 25Hz for Liver/WAT or for 5 minutes @ 30Hz for muscle
#Homogenize by sonicating on ice 3 x 15s or 3 x freeze thaws.
#Add 12.5ul KOH
#Add 5 ul KOH
#Mix by inverting  
#Mix by inverting  
#Add 800ul '''Chloroform/Methanol Mixture'''
#Add 800ul '''Chloroform/Methanol Mixture'''