Confocal immunofluorescent analysis of HeLa cells using MitoTracker® Red CMXRos (red) and Lamin A/C (4C11) Mouse mAb (Alexa Fluor® 488 Conjugate) #8617 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Each vial of MitoTracker® Red CMXRos contains 50 μg lyophilized solid. To make a 1 mM stock solution, reconstitute the solid in 94.1 μl high quality DMSO. While the optimal staining concentration and incubation times can vary by application, typical results are obtained by diluting the stock solution directly into growth media at a concentration between 50-200 nM and incubation between 15-45 min at 37ºC. After incubation, fix the cells in ice-cold methanol for 15 min at -20ºC then rinse 3 times with PBS for 5 min. Proceed with immunostaining.
Store lyophilized solid at -20°C dessicated and protected from light. In lyophilized form, this reagent is stable for 12 months. Once reconstituted in DMSO, the solution can be stored at -20°C, protected from light and must be used within 2 weeks. Avoid freeze-thaw cycles.
MitoTracker® Red CMXRos is well retained after fixation allowing for further sample processing and immunostaining. Excitation: 579 nm, Emission: 599 nm, Molecular Weight: 531.52
MitoTracker® dyes are cell permeable probes that contain a mildly thiol-reactive chloromethyl moiety for mitochondrial labeling. To label mitochondria, cells are simply incubated with MitoTracker® probes, which passively diffuse across the plasma membrane and accumulate in active mitochondria.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
DRAQ5 is a registered trademark of Biostatus Limited.
MitoTracker is a registered trademark of Molecular Probes, Inc.