I bought this Alcohol Dehydrogenase inhibitor awhile back and foolishly did not aliquot out portions of the inhibitor before storing it long time. The original vial was stored at 4C (per instructions) and has been through probably 4 or 5 cycles of "freeze/thaw." I suspect the inhibitor is sensitive to this kind of treatment and thus the inhibitor is now bad. Can anyone give me an insight to what I suspect?
I'm currently assessing whether or not the inhibitor works in flies by assessing their activity during ethanol vapor exposure. I used the inhibitor a few months back an was seeing modest effects in the response to ethanol. Now I am not seeing anything.
Thanks RGC
The flashpoint of 4-methylpyrazole is 93 degrees celsius so you might have to be careful about it if your ADH are Extremophile (Thermophile) derived enzymes. But if your concern is related to storage then my question is, has the chemical darkened? If so, there is still no need to be concerned as it 4-methylpyrazole doesn't lose its purity. Freshly prepared one would be colourless to yellow and it does darken after prolonged storage, but the properties of the chemical doesn't change. Unless your dehydrogenase is derived from a thermophilic organism like Sulfolobus, where it's probably best to find the specific activity at various temperatures and then back calculate the inhibitory mechanism. For ex: If the enzyme is active at 37 degrees at 55% the activity from the optimum temperature (say 70 degrees) then inhibitory mechanism can be back-calculate. But it would be an approximate estimation.
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