I have seen a protocol suggesting using adenosine deaminase to transform adenosine to xanthine in deproteinized media and then measure xanthine, but no details were provided.
I have also been across papers where ATP or cAMP is measured instead.
Which method is more accurate?
Thank you in advance!
Depending on the amount you may use just commercial adenosine deaminase and measure the decrease
in absorbance in a fluorimeter. You need a source of UV light and appropriate cuvettes (transparent to UV light), and I would think that 265 nm was the right wavelength. If needed I can look for one of our "Old" papers when me measured enzyme activity (and send it to you).
If you have an estimation of adensoine concentrations you may check whether the method is sensitive enough. Probably it is sensitive enough.
The method was standard and can be used the other way around, i.e. instead of measuring how much adenosine deaminase, you may use it to measure how much adenosine.
Important you can probably use this method in culture supernantants not in extract from cells as Ado would be quickly converted into other "things"!!!. And finally, measurements should be done quickly just in case adenosine is being transformed by the presence of enzymes in the culture medium. If you measure quick, the enzymes in the clture medium would not change much concentration but if you wait and wait, adenosine may be transformed to other compounds.
Dear Rafael, thank you for your helpful answer. I would be grateful if you could send me one of your "old" papers.
Too old to get their pdfs from databases. I will retrive the paper(s) from my files and I will do a pdf for one or two of my articles . If there is delay please remind me on Wednesday (tomorrow Tues. I have a difficult day)
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