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. Author manuscript; available in PMC: 2023 Sep 12.
Published in final edited form as: Methods Enzymol. 2020 Jun 13;641:75–94. doi: 10.1016/bs.mie.2020.04.037

Fig. 2.

Fig. 2

Overview of IMPACT with 3-azidopropanol and SPAAC detection. (A) Workflow for IMPACT labeling and detection of PLD activity by flow cytometry, HPLC with fluorescence detection, and mass spectrometry. (B) Flow cytometry histogram of HeLa cells labeled using 3-azidopropanol and BCN-BODIPY in IMPACT. (C) HPLC chromatograph of a lipid extract of HeLa cells after IMPACT labeling with 3-azidopropanol and click chemistry tagging of lipid extracts with BCN-BODIPY. (D) Quantification of the phosphatidyl alcohol products generated when HeLa cells were treated with 3-azidopropanol and PLD activity was stimulated by PMA. Lipid extracts were reacted via click chemistry with Alk-QA to increase detection sensitivity. This figure was adapted with permission from ACS Central Science (https://pubs.acs.org/doi/abs/10.1021/acscentsci.7b00222). Further permissions requests for this material should be directed to the ACS.