To determine the NO scavenging acitivty of plant extracts, I am following (Sreejayan & Rao, 1997) protocol. I could not able to produce reproducible results.

1.Is that we can start experiment after preparing 5mM SNP or we have to wait for particular time (Say 6 hours)?

2 . I read in one reference the Light is very important to perform the reaction. So, I am doing this experiment closed room under 40 watts Tube light ( A fluorescent lamp, or fluorescent tube, is a low-pressure mercury-vapor gas-discharge lamp that uses fluorescence to produce visible light ).

3. In Griess reagent (1% sulphanilamide, 2% H3P04 and 0.1% naphthy- lethylenediamine dihydrochloride), can we directly dissolve in PBS buffer ?

Please Suggest me.

Thank you.