Ideally I would like to be able to image live and also to be able to make fixed slides. I want to probe for the cellular distribution of the variously phosphorylated phosphatidylinositols.
I am trying to go through various companies no luck so far.
Hi Neal Edward Beeman . Avanti Polar Lipids has high quality lipid products including PI and fluorescent PI.
TopFluor® Derivatives (avantilipids.com)
Echelon Biosciences
https://echelon-inc.com
also has a range of PI lipids, antibodies and protein binding domains that distinguish between the different phosphoinositides.
There are various PIns-binding motifs fused to GFP that can be used to monitor different phosphoinositide species such as PH-domain of PLC (GFP- PH-PLC d) can be used for PI(4,5)P2, PH-GFP AKT for PI(3,4,5)P3, PH-OSBP (PI4P) etc.
Article Visualization of Cellular Phosphoinositide Pools with GFP-Fu...
doi: 10.1152/physrev.00028.2012
I'd recommend checking out some of the work that Gerry (GRV) Hammond has done imaging these lipids. He's been involved with some lovely studies.
for example
Article Probing the subcellular distribution of phosphatidylinositol...
and
https://pubmed.ncbi.nlm.nih.gov/22722250/
My lab mainly uses the lipid binding domains for live cell imaging
The antibodies I've mainly used for semi-quantifying extracted lipids in lipid blots
Great answers.
I went to addgene and was able to get several of the constructs suggested here
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