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The Histoblot Technique: A Reliable Approach to Analyze Expression Profile of Proteins and to Predict Their Molecular Association

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Co-Immunoprecipitation Methods for Brain Tissue

Part of the book series: Neuromethods ((NM,volume 144))

Abstract

The histoblot technique, a hybrid of immunohistochemistry and immunoblotting techniques, has emerged as an attractive alternative to study the expression levels of proteins. This method is an easy, fast, and widely used approach in neuroanatomy that provides a reliable way to reveal and compare the regional distribution and expression profile of different proteins in the brain. The histoblot approach does not require the use of fixatives, which are employed for conventional immunohistochemistry. This avoids chemical modifications that altering the integrity of antibody-binding sites can make difficult the access of antibody to epitopes. Instead, the histoblot technique requires similar experimental conditions to those of immunoblot analysis of protein extracts from dissected brain regions, except that for histoblots tissue samples are not homogenized and protein transfer is achieved by mechanical pressure. Therefore, the histoblot reflects the spatial pattern in which proteins are arranged within a brain section, rather than protein bands, providing more accurate and direct information on the anatomical localization of proteins. Furthermore, though histoblot is essentially a macroanatomic technique, it exhibits a high subregional resolution. Based on comparative analyses between different proteins, the histoblots also help to predict their possible molecular interaction. However, the main disadvantage is that this method is not suitable to unravel protein distribution at the cellular and subcellular levels. In this chapter, we introduce the histoblot procedure used primarily on brain sections for the identification of quantitative changes of neurotransmitter receptors and ion channels and also discuss the limitations inherent to these approaches.

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Acknowledgments

This work was supported by the grants BFU2015-63769-R from the Spanish Ministry of Education and Science, the European Union (HBP - Project Ref. 720270), and the Junta de Comunidades de Castilla-La Mancha (SBPLY/17/180501/000229).

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Authors and Affiliations

  1. Facultad de Medicina, Departamento de Ciencias Médicas, Instituto de Investigación en Discapacidades Neurológicas (IDINE), Universidad Castilla-La Mancha, Albacete, Spain

    Carolina Aguado & Rafael Luján

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  1. Carolina Aguado
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  2. Rafael Luján
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Correspondence to Rafael Luján .

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Editors and Affiliations

  1. Department of Psychiatry, Saitama Medical University, Saitama, Japan

    Yuji Odagaki

  2. Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden

    Dasiel O. Borroto-Escuela

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Aguado, C., Luján, R. (2019). The Histoblot Technique: A Reliable Approach to Analyze Expression Profile of Proteins and to Predict Their Molecular Association. In: Odagaki, Y., Borroto-Escuela, D. (eds) Co-Immunoprecipitation Methods for Brain Tissue . Neuromethods, vol 144. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8985-0_6

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  • DOI: https://doi.org/10.1007/978-1-4939-8985-0_6

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8984-3

  • Online ISBN: 978-1-4939-8985-0

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