Are there any non enzymatic methods to determine IP3 generation other than radio-tagged method?

11 November 2013 1 4K Report

We are looking to quantify IP3 generation through in vitro method. We need a simple analytical method to measure it.

I've been following this question for a while, hoping someone would know. Unfortunately, I don't have an answer, I've been trying to find something for a few years and people in Cell Bio that I've asked have told me that basically IP3 is difficult to measure with any consistency.

The one thing I've found (that I will try soon) isn't very simple, but we have the setup for it and it looks interesting: a FRET reporter of PIP2 cleavage activity called 'CYPHR' (at Addgene.org, Plasmid 14864, which will link to the publication that discusses it). This isn't exactly in vitro, but just mentioning it in case it can fit your needs.

Good luck!

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