Dear Abdalla, Thank you so much for your answer, but it is an inhibitor for LPL too?
best
Mehdi
Dear Apurva, Dear Abdalla, Thank you very much for your kind replying. But my purpose is enzyme inhibition in vitro (in test tube), e,g I found that NaCl 1M can inhibit the LPL more than 50% in vitro. I am looking for an more effective inhibitor (s) with lower concentration and higher inhibition.
best
mehdi
Oh OK I got it. I think u wish to inhibit lpl while leaving other esterase activities intact. Is that so?
Sir, lpl needs apocII , which is incorporated in to assay by addition of heat inactivated serum. Elimination of this step could ensure reduced lpl activity which could be further taken care of by nacl. Will this work for you?
Dr. El-Mowafy, what you say is absolutely correct. Orlistat is almost non specific.
Orlistat inhibits LPL (s. Eur J Biochem. 1994 Jun 1;222(2):395-403) and you can buy it from Sigma, so you don´t have to resolve a pill. But as mentioned above by others orlistat inhibits other lipases, too.
Inhibition by 1M NaCl is an often used method but for some reasons its not always working completely. Seems to be that it is dependent on the used substrate in the activity test. If you are working with the commercial kits based on a fluorescence omittance of apoCII doesn´t work because in these test LPL don´t need the cofactor.
Dear Michael,
Thank you for your helpful answer.
I have a mixture of three enzymes in my sample, LPL, GK(Glycerol kinase), and GPO (Glycerol phosphate oxidase). substrate is TG (triacylglycerol), I need inhibition of LPL in different time and measuring Glycerol using another two enzymes in the media.
What is your suggestion for Orlistat concentration?
e.g. I have 5 test tubes contain 2kU/l LPL, and 0.5 Ku/l GK and 0.5 KU/L GPO.
of course and Mg++, ATP, Buffer,...
I want to add TG to all test tubes at 0 time and after 2min stop the LPL reaction in first test tube, after 4min stop the LPL reaction in the second test tube and so on.
best
mehdi
The authors of the following article used 1 µM (they solved it in ethanol)
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4284735/
I am of the same opinion as that of Dr. Michael Marcus Hoffmann.
- Badges
- Science topic
- Similar topics
- Biological Science
- Microbiology
- Virology
- Virus