Skip to main content

Immunohistochemical Detection of Alpha-Synuclein in Unfixed Human Brain Tissue

  • Protocol
  • First Online:
Alpha-Synuclein

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1948))

Abstract

In both research and diagnostics, immunohistochemistry is an essential method for assessing pathology in neurodegenerative diseases. Typically, at autopsy, one hemisphere of the brain is formalin fixed for sectioning and histochemical analysis, while the other hemisphere is flash frozen for biochemical studies. While this process ensures the generation of high-quality slides, it is important to assess the pathology of tissue near pieces used for biochemical analyses, since pathology burden can be unevenly distributed throughout the brain (Fig. 1). Here, we describe a method for assessing alpha-synuclein (αSyn) pathology in unfixed human brain tissue using a novel monoclonal antibody.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 169.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Reiner L (1930) On the chemical alteration of purified antibody-proteins. Science 72(1871):483–484

    Article  CAS  Google Scholar 

  2. Marrack J (1934) Nature of antibodies. Nature 133:292–293

    Article  CAS  Google Scholar 

  3. Coons AH, Creech HJ, Jones RN (1941) Immunological properties of an antibody containing a fluorescent group. Exp Biol Med 47(2):200–202

    Article  CAS  Google Scholar 

  4. Graham RC, Lundholm U, Karnovsky M (1965) Cytochemical demonstration of peroxidase activity with 3-amino-9-ethylcarbazole. J Histochem Cytochem 13:150–152

    Article  CAS  Google Scholar 

  5. Graham RC, Karnovsky MJ (1965) The histochemical demonstration of uricase activity. J Histochem Cytochem 13(6):448–453

    Article  CAS  Google Scholar 

  6. Graham RC, Karnovsky MJ (1965) The histochemical demonstration of monoamine oxidase activity by coupled peroxidatic oxidation. J Histochem Cytochem 13(7):604–605

    Article  CAS  Google Scholar 

  7. Avrameas S, Uriel J (1966) Method of antigen and antibody labelling with enzymes and its immunodiffusion application. C R Acad Sci Hebd Seances Acad Sci D 262(24):2543–2545. French

    CAS  PubMed  Google Scholar 

  8. Nakane PK, Pierce GB (1966) Enzyme-labeled antibodies: preparation and application for the localization of antigens. J Histochem Cytochem 14(12):929–931

    Article  CAS  Google Scholar 

  9. Childs, G (2014) History of immunohistochemistry. In: McManus LM, Mitchell RN (ed) Pathobiology of human disease, 1. Academic Press, Cambridge, MA

    Google Scholar 

Download references

Acknowledgments

The author would like to thank Dr. Mel Feany, Dr. Liqun Wang, and Kit Sin Tuen from the Feany Laboratory at the Ann Romney Center for Neurologic Disease at Brigham and Women’s Hospital for assistance with developing the protocol. The author would also like to acknowledge the Pathology Core at the Dana-Farber Cancer Institute for their work in optimizing the 2F12 antibody concentration.

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2019 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Sanderson, J.B. (2019). Immunohistochemical Detection of Alpha-Synuclein in Unfixed Human Brain Tissue. In: Bartels, T. (eds) Alpha-Synuclein. Methods in Molecular Biology, vol 1948. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9124-2_2

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-9124-2_2

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9123-5

  • Online ISBN: 978-1-4939-9124-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics