Immunohistochemical Detection of Alpha-Synuclein in Unfixed Human Brain Tissue

  • John B. Sanderson
Part of the Methods in Molecular Biology book series (MIMB, volume 1948)


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.

Key words

Alpha-synuclein Immunohistochemistry Human tissue Frozen Fresh 



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.


  1. 1.
    Reiner L (1930) On the chemical alteration of purified antibody-proteins. Science 72(1871):483–484CrossRefGoogle Scholar
  2. 2.
    Marrack J (1934) Nature of antibodies. Nature 133:292–293CrossRefGoogle Scholar
  3. 3.
    Coons AH, Creech HJ, Jones RN (1941) Immunological properties of an antibody containing a fluorescent group. Exp Biol Med 47(2):200–202CrossRefGoogle Scholar
  4. 4.
    Graham RC, Lundholm U, Karnovsky M (1965) Cytochemical demonstration of peroxidase activity with 3-amino-9-ethylcarbazole. J Histochem Cytochem 13:150–152CrossRefGoogle Scholar
  5. 5.
    Graham RC, Karnovsky MJ (1965) The histochemical demonstration of uricase activity. J Histochem Cytochem 13(6):448–453CrossRefGoogle Scholar
  6. 6.
    Graham RC, Karnovsky MJ (1965) The histochemical demonstration of monoamine oxidase activity by coupled peroxidatic oxidation. J Histochem Cytochem 13(7):604–605CrossRefGoogle Scholar
  7. 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. FrenchPubMedGoogle Scholar
  8. 8.
    Nakane PK, Pierce GB (1966) Enzyme-labeled antibodies: preparation and application for the localization of antigens. J Histochem Cytochem 14(12):929–931CrossRefGoogle Scholar
  9. 9.
    Childs, G (2014) History of immunohistochemistry. In: McManus LM, Mitchell RN (ed) Pathobiology of human disease, 1. Academic Press, Cambridge, MAGoogle Scholar

Copyright information

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

Authors and Affiliations

  • John B. Sanderson
    • 1
  1. 1.The Warren Alpert Medical School of Brown UniversityProvidenceUSA

Personalised recommendations