I need to know hoe we can calculate how much induction of inos is there in treated compared to control with absorbance value. Hoe to use standard curve value in exp values??
The Griess reaction doesn't tell you how much inducible nitric oxide synthase activity there is. It just measures nitrite concentrations. In biological samples these may come from NOS1-3 (i.e. endothelial, inducible, neuronal as they used to say).
You'll need to construct a standard curve of (usually sodium or potassium) nitrite, then read off your samples.
Make sure it's nitrIte, not nitrAte.
Hello, Simon and Daniel are right. When you measure the NO by using Griess Assay, it will not tell you the iNOS level. If you want to know the iNOS level, I will recommend you to detect the iNOS mRNA and protein level, using Realtime-PCR and WB, respectively. As I found before, the NO amount in Griess Assay is varied by the cell amount you cultured or seed. So, to confirm the results, we often detect iNOS mRNA level as well as NO level by Griess Assay.
Hope to help you, good luck
- Thanks Lee, earlier I use to check with inos primer only but it's not showing induction although upregulation of il-beta is there.actually I.am infecting raw cells with H.pylori.thereafter my guide suggest me to use griess reagent but even with upregulation of 95 fold up il-beta,there is no. Olor change during inos detection.
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