How to estimate intracellular metabolite concentration?

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Pankaj Thapa

Have you considered relying on NMR for addressing similar kind of question? I mean, NMR metabolite profiling using 1D NMR is one of the recent advances which researcher are using.

Jai Ghosh

I usually do it by rapidly freezing the cells to-70 or-80C and then homogenize it at that temperature usually by ultrasonication. Carry out the extraction depending upon the metabolite you are looking for and check it say by MS.

Inamullah Hakeem Said

I would second Mr.Jai Ghosh's answer. You can follow the extraction protocol in section 2.6 in one of my paper. You can modify it and may help you.

Article Quantification of microbial uptake of quercetin and its deri...

Mr. Thapa: 1D NMR for metabolite profiling is no doubt a very handy tool. However, It is most cumbersome for intermediate metabolites especially when these are intra cellular.

Rushikesh Patil

Thank you very much for sharing your opinion. It is helpful. What will be the internal standards for glycolytic and TCA cycle metabolites estimation to be mixed with metabolite cellular extract ? Are they commercially available?