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AAT Bioquest

Is mitochondria stain, MitoLite™ Red CMXRos fixable after staining?

Posted October 21, 2019


Answer

Yes, MitoLite™ RED CMXRos stain is well-retained after fixation.

To stain mitochondria of live cells, prepare a 100–500 nM working solution of MitoLite™ Red CMXRos. After staining live cells with MitoLite™ Red CMXRos, cells can be fixed and permeabilized for subsequent probing/staining of other intracellular structures by immunocytochemistry (ICC).

Fixation and permeabilization protocol

  1. After staining, wash cells in fresh, pre-warmed buffer or growth medium.
  2. Remove buffer or medium, and replace with 3.7% formaldehyde in complete growh medium at 37°C for 15 minutes.
  3. After fixation, rinse cells several times in buffer.
  4. Permeabilization (optional): Incubate fixed cells in buffer containing 0.2% Triton® X-100 for 10 minutes. Following permeabilization, rinse cells in buffer before proceeding with any subsequent ICC staining.
Additional resources

MitoLite™ RED CMXRos