I am determining the lipid profile of fish liver and muscle by simply making the homogenate in phosphate buffer (50mM, ph 7.2) centrifuge and then proceed according to the protocol given in the estimation kits for serum. Is that OK, or do I first need to extract the lipid from tissue and then proceed with the diagnostic kits?
This phosphate buffer which is named PBS{ Phosphate-Buffered Saline;7.2 pH} is one of the most common buffer used in biochemistry during cell structure elucidation by a spectroscopic technique called ESR( Electron Spin Resonance).
All you have to do is to add lipid in PBS and mix the two thoroughly and take the reading.
“The phosphate buffer is HYDROPHILIC and the lipids ARE ALSO PHOSPHATE BASED LIPOPHILIC substances. In addition, this buffer is ISOTONIC{ same osmotic pressure) and NON-TOXIC to most cells and thus the cell structure does not change during the conduct of the experiment.
Which kits will you use? There are several kits like Infinity for TAG or the NEFA kit for free fatty acids. Especially when you use the Infinity TAG kit you have to consider that this kit totally hydrolyzes TAG, DAG and MAG which is no problem in serum but can give you big errors in liver.
The homogenate should be ok for this kind of determination. All chemicals which are not compatible with the kit, are usually listed in the manual.
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