TLC spots in enzyme assays of lipids

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hibio
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TLC spots in enzyme assays of lipids

hi all,
If one does MGAT and DGAT assays, why not only the formation of DAG and TAG some other spots will come in the TLC separated. Wt might be the reason for this?
 

R Bishop
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hibio,

hibio,
Can you clarify?  Are you asking why secondary TLC spots appear other than diacylglycerol and triacylglycerol appear on the plate?
 
Rus

hibio
hibio's picture
hi

hi

Yes my question is why one see secondary spots. More cleary, for an example in DGAT assay with purified DGAT protein (not the crude cytosolic fraction), we are using DAG and labeled fattyacids are using as substrates. After the reaction, extract lipids and run on TLC, why do we see Free fattyacids, MAG also DAG (all are labelled) apart from only TAG. What might be the reasong. not only that, if u do time dependent assay these spots also shows increase along with TAG.

Thanx for reply
hibio