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Procedures for ADC Immunoblotting and Immunolocalization for Transmission Electron Microscopy During Organogenic Nodule Formation in Hop

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Polyamines

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1694))

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Abstract

Immunolocalization for transmission electron microscopy is a powerful technique to identify subcellular localization of proteins. This can be combined with molecular and physiological data in order to have a complete overview of protein function. However, optimal sample preservation is required to avoid artefacts. When using chemically fixed samples, the progressive lowering of temperature (PLT) technique is a convenient procedure to dehydrate and embed samples at low temperature, thereby preserving the antigenicity of the proteins to be detected. Despite the advantages of immunogold labelling, it is a time-consuming cell biology technique. Therefore, the quality and specificity of the antibody should be previously checked by western blot. This approach also enables to identify changes in the amount of protein under study throughout development or in response to stress conditions.

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Acknowledgments

This work was supported by the grant AGL2014-55177-R from Spanish MINECO to JMSS. Funding to AMF was provided by the Portuguese Foundation for Science and Technology (SFRH/BPD/100928/2014, FCTInvestigator IF/00169/2015, PEst-OE/BIA/UI4046/2014. We also acknowledge The Society of Plant Signaling and Behavior for allowing reproduction of images.

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Authors and Affiliations

  1. Biosystems & Integrative Sciences Institute (BioISI), Faculdade de Ciências, Universidade de Lisboa, Campo Grande, 1749-016, Lisbon, Portugal

    Ana Margarida Fortes

  2. Cell Biology Group–COMAV, Universitat Politècnica de València, CPI, Edificio, 46022, Valencia, Spain

    Jose M. Seguí-Simarro

Authors
  1. Ana Margarida Fortes
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  2. Jose M. Seguí-Simarro
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Correspondence to Ana Margarida Fortes .

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Editors and Affiliations

  1. Department of Biology, Healthcare and Environment, Faculty of Pharmacy and Food Sciences, Section of Plant Physiology, University of Barcelona, Spain

    Rubén Alcázar

  2. Department of Biology, Healthcare and Environment, Faculty of Pharmacy and Food Sciences, Section of Plant Physiology, University of Barcelona, Spain

    Antonio F. Tiburcio

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Fortes, A.M., Seguí-Simarro, J.M. (2018). Procedures for ADC Immunoblotting and Immunolocalization for Transmission Electron Microscopy During Organogenic Nodule Formation in Hop. In: Alcázar, R., Tiburcio, A. (eds) Polyamines. Methods in Molecular Biology, vol 1694. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7398-9_19

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  • DOI: https://doi.org/10.1007/978-1-4939-7398-9_19

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7397-2

  • Online ISBN: 978-1-4939-7398-9

  • eBook Packages: Springer Protocols

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