I want to perform an inhibition study of iNOS activity in primary hepatocyte by using aminoguanidine. What concentration of aminoguanidine in cell culture should I choose?
Thanks in advance.
Hi
Look this link PLZ
Effects of a nitric oxide donor and nitric oxide synthase inhibitors on ...
humrep.oxfordjournals.org/content/14/10/2537.fullHuman Reproduction
by K Mitsube - 1999 - Cited by 31 - Related articles
In studies using cell culture models, NO consistently inhibits oestradiol and .... The results of ovulation number, concentrations of steroids and NO products at ... the induction of inducible nitric oxide synthase ( iNOS) in rat ovaries perfused for 10 h ... Administration of NOS inhibitors (aminoguanidine bicarbonate, AG 1 mmol/l ...
I wouldn't use aminoguanidine: it is not selective for iNOS vs several other targets, especially at 1mM!
There are several better inhibitors, e.g. 1400W: http://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Product_Information_Sheet/1/w4262pis.pdf
Note that these inhibitors are usually competitive with arginine so you need higher concentrations at higher arginine.
Good luck!
Hi Richard,
I am looking for NOS inhibitors, not selective, but able to inhibit all isoforms. When you say Aminoguanidine is not selective for iNOS, does it mean it can inhibit other isoforms too? Basically my question is if there is any inhibitor which can inhibit all isoforms of NOS. I know that the efficiency is going to be different. Do you think that using a substrate analogue like L-NNA, or L-NAME is good since most of the NOS isoforms have highly similar active sites?
Hi Sanjay,
Both L-NNA or L-NAME should work. There are several NOS inhibitors, you may use but IC50 varies. So you should think about the concentration you are going to use.
Here is a good reference for you,
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1221991/pdf/11463332.pdf
Hi Abdul,
Thank you for your response. Probably I will calculate the IC50 for both L-NNA or L-NAME using cells of my interest and then use both of these analogues for my assay.
Best wishes,
Sanjay
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