Recipe

Staining mix B

Reagent Amount to add (for 20 mL) Final concentration
Dulbecco’s PBS for lipid labeling 19.975 mL
3-Azido-7-(diethylamino) coumarin (40 mM prepared in DMSO)a 25 μL 50 μM
Prepare the mix fresh immediately before each labeling experiment.
aSee Sivakumar et al. 2004 (Sivakumar K, Xie F, Cash BM, Long S, Barnhill HN, Wang Q. 2004. A fluorogenic 1,3-dipolar cycloaddition reaction of 3-azidocoumarins and acetylenes. Org Lett 6: 4603-4606).
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  1. doi:10.1101/pdb.rec12262 Cold Spring Harb Protoc 2010: pdb.rec12262-
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