Abstract
Immunofluorescence staining is a widely used and powerful tool for the visualization and colocalization of two or more proteins and/or cellular organelles. For colocalization studies in fixed cells, one target protein/organelle is immunostained and visualized by one fluorophore and the other target protein/organelle is immunostained and visualized by a different fluorophore whose excitation emission spectra does not overlap with the first fluorophore. Parkin (PARK2) is an E3 ubiquitin ligase which performs ubiquitination of surface proteins of dysfunctional mitochondria to mark them for autolysosomal degradation. Here we describe the immunofluorescence staining of parkin protein and immunofluorescence or dye-based methods to visualize mitochondria and study the colocalization of parkin and mitochondria in primary human or mouse chondrocytes or cell lines.
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Acknowledgments
This work was supported by grants from National Institute of Arthritis and Musculoskeletal and Skin Diseases and National Center for Complementary and Integrative Health of the National Institutes of Health under award numbers R01AR067056, AT007373, and by funds from Northeast Ohio Medical University to TMH.
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Ansari, M.Y., Haqqi, T.M. (2021). Assessing Chondrocyte Status by Immunofluorescence-Mediated Localization of Parkin Relative to Mitochondria. In: Haqqi, T.M., Lefebvre, V. (eds) Chondrocytes. Methods in Molecular Biology, vol 2245. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1119-7_15
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DOI: https://doi.org/10.1007/978-1-0716-1119-7_15
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