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ID 114466
Title Alternative
Sensitive immunohistochemistry for autopsied brain
Author
Keywords
immunohistochemistry
polymer staining
tyramide signal amplification
avidin biotin complex
humans
brain
striatum
neuroanatomy
Content Type
Journal Article
Description
Immunohistochemistry (IHC) is a valuable method for identifying discrete neurochemical molecules by the interaction of target antigens with validated antibodies tagged with a visible label (e.g., peroxidase). We have developed an immunostaining method that is highly sensitive in detection of neurochemical antigens. Our IHC method, which we call the PBTA method, involves a hybrid protocol that implements aspects of both the polymer and avidin-biotin-complex (ABC) methods in combination with biotin-tyramide amplification. When using [Met]-enkephalin as a target antigen, the sensitivity of the PBTA method for IHC was more than 100-fold higher compared with the polymer and ABC methods. In addition, its sensitivity for enzyme-linked immunosorbent assay was about 1,000-fold higher compared with the ABC method. We examined the utility of our IHC method for both chromogenic and fluorescence detection systems used to visualize neurochemical peptides and proteins in formalin-fixed, paraffin-embedded tissues from autopsied human brains. The results convincingly demonstrate that under optimal conditions, our IHC method is highly sensitive without increasing non-specific background activities. Our IHC method could be a powerful tool for detection and visualization of neurochemical antigens that are present even in trace amounts in autopsied human brains.
Journal Title
Frontiers in Neuroanatomy
ISSN
16625129
Publisher
Frontiers Media S.A.
Volume
9
Start Page
22
Published Date
2015-03-03
Rights
© 2015 Goto, Morigaki, Okita, Nagahiro and Kaji. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY)(https://creativecommons.org/licenses/by/4.0/). The use, distribution and reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
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language
eng
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departments
Medical Sciences
University Hospital