LysoSensor PDMPO

Guide (The following is a recommended protocol. This protocol is provided as a guide only and should be modified to suit your specific needs).
1. Preparation of Stock Solution^[1]
1.1 Dissolve LysoSensor PDMPO in 50 μL of anhydrous DMSO at a concentration of 1 mM to prepare the stock solution.
1.2 Aliquot the stock solution to avoid repeated freeze-thaw cycles.
2. Preparation of Working Solution
2.1 Dilute the 1 mM stock solution to a working concentration of 1-200 μM in antibiotic-free DMEM medium (the recommended initial concentration is 1 μM).
Note: Prepare the working solution freshly before use. Thaw the aliquoted stock solution at room temperature prior to use and protect from light.
3. Staining Procedure
3.1 Fixed Cell Imaging
3.1.1 Incubate the cells with LysoSensor PDMPO working solution at 37°C and 5% CO₂ for 5 min.
3.1.2 Discard the staining solution and wash the cells 3 times with pre-chilled PBS to remove unbound dye.
3.1.3 Fix the cells with 4% PFA at room temperature in the dark for 15 min.
3.1.4 Discard the PFA and wash the cells 3 times with PBS.
3.1.5 Imaging detection.
3.2 Live Cell Imaging
3.2.1 Incubate the cells with LysoSensor PDMPO working solution at 37°C and 5% CO₂ for 5 min.
3.2.2 Discard the staining solution and gently wash the cells 3 times with PBS.
3.2.3 Place the cells in a live-cell imaging system, and acquire images at a rate of 1 frame/30 s for 1 h.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.