Organelle isolation from mouse tissues expressing organelle tags

Output Details

We describe here a method to perform the rapid isolation of intact lysosomes from mouse tissues expressing a lysosome-localized TMEM192-3×HA fusion protein (“LysoTag”) (as in JAX stock #035401, RRID:IMSR_JAX:035401). The fusion protein marks lysosomes with a triple HA epitope, enabling the rapid isolation of intact tagged organelles from tissues using an anti-HA antibody. The organelles purified using this method are highly enriched, intact, largely contaminant-free and can be used for various downstream applications, including immunoblotting analysis and proteomic analysis (as described in dx.doi.org/10.17504/protocols.io.ewov1o627lr2/v1), but also lipidomic or metabolomic analysis (as described in dx.doi.org/10.17504/protocols.io.bybjpskn).
Tags
  • Lysosomes

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