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Step-by-Step Protocol for Superparamagnetic Nanoparticle-Based Endosome and Lysosome Isolation from Eukaryotic Cell

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Stem Cell Nanotechnology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2125))

Abstract

Here, we report our step-by-step protocol for superparamagnetic nanoparticle (SPMNP)-based endosome and lysosome isolation from HeLa. Briefly, we synthesized SPMNP 1.0 with iron oxide (Fe3O4) core using thermal decomposition method. Further, we performed ligand-exchange strategy for surface functionalization of SPMNP 1.0 with dimercaptosuccinic acid (DMSA). Thus, we generated DMSA-SPMNP 2.0 and used DMSA-SPMNP 2.0 to isolate endosomes and lysosome from HeLa cells. Using our SPMNP subcellular fractionation protocol, we are able to isolate high-pure-high-yield lysosomes using DMSA-SPMNP 2.0 for lysosome proteomics and lipidomics in order to better understand subcellular compartments.

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References

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Acknowledgments

This work was supported by Envirotransgene® Global. Authors thank the infrastructural support from Bannari Amman Institute of Technology, India.

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Authors and Affiliations

  1. Envirotransgene® Bio-solutions Global Chennai India and Institute of Cancer Research, Oslo University Hospital, Oslo, Norway

    Deepak B. Thimiri Govinda Raj

  2. Laboratory of Lipid Metabolism and Cancer, Louvain, Belgium

    Niamat Ali Khan

  3. Bannari Amman Institute of Technology, Sathyamangalam, TN, India

    Srisaran Venkatachalam

  4. Faculty of Biology, Hanoi National University of Education, Hanoi, Vietnam

    Dinh Toi Chu

  5. Institute for Research and Development, Duy Tan University, Danang, Vietnam

    Dinh Toi Chu

  6. School of Bio Sciences and Technology (SBST), VIT University, Vellore, TN, India

    Sivakumar Arumugam

  7. School of Odonto Stomatology, Hanoi Medical University, Hanoi, Vietnam

    Dinh Toi Chu

Authors
  1. Deepak B. Thimiri Govinda Raj
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  2. Niamat Ali Khan
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  3. Srisaran Venkatachalam
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  4. Dinh Toi Chu
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  5. Sivakumar Arumugam
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Corresponding author

Correspondence to Deepak B. Thimiri Govinda Raj .

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Editors and Affiliations

  1. Ottawa, ON, Canada

    Kursad Turksen

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Thimiri Govinda Raj, D.B., Khan, N.A., Venkatachalam, S., Chu, D.T., Arumugam, S. (2019). Step-by-Step Protocol for Superparamagnetic Nanoparticle-Based Endosome and Lysosome Isolation from Eukaryotic Cell. In: Turksen, K. (eds) Stem Cell Nanotechnology. Methods in Molecular Biology, vol 2125. Humana, New York, NY. https://doi.org/10.1007/7651_2019_212

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  • DOI: https://doi.org/10.1007/7651_2019_212

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0359-8

  • Online ISBN: 978-1-0716-0360-4

  • eBook Packages: Springer Protocols

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