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Plant Cytogenetics pp 161–175Cite as

Immunolocalization on Whole Anther Chromosome Spreads for Male Meiosis

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1429))

Abstract

Immunolocalization of cells undergoing meiosis has proven to be one of the most important tools to decipher chromatin-associated protein dynamics and causal relationships. Here, we describe a protocol established for maize which is easily adaptable to other plants, for example, with minor modifications to Arabidopsis as stated here. In contrast to many other protocols, the following protocol is based on fixation by a 3:1 mixture of ethanol and acetic acid. Spreading of cells is followed by freeze-shattering, protein antigenicity retrieval by a hot citrate buffer bath, antibody incubations and washes, and DNA staining.

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Acknowledgments

We are grateful to Drs. Zac Cande, Wojciech Pawlowski, and Rachel Wang for the maize ASY1 and ZYP1 antisera, and to Chris Lambing for helpful discussion on Fig. 4. This work was supported by the National Science Foundation (IOS: 1025881).

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Authors and Affiliations

  1. Department of Horticultural Science, University of Minnesota, Alderman Hall, 1970 Folwell Ave., St. Paul, MN, USA

    Stefanie Dukowic-Schulze, Anthony Harris & Changbin Chen

Authors
  1. Stefanie Dukowic-Schulze
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  2. Anthony Harris
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  3. Changbin Chen
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Corresponding author

Correspondence to Changbin Chen .

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Editors and Affiliations

  1. University of Minnesota, St. Paul, Minnesota, USA

    Shahryar F. Kianian

  2. University of Minnesota, St. Paul, Minnesota, USA

    Penny M. Avoles Kianian

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Dukowic-Schulze, S., Harris, A., Chen, C. (2016). Immunolocalization on Whole Anther Chromosome Spreads for Male Meiosis. In: Kianian, S., Kianian, P. (eds) Plant Cytogenetics. Methods in Molecular Biology, vol 1429. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3622-9_13

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  • DOI: https://doi.org/10.1007/978-1-4939-3622-9_13

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3620-5

  • Online ISBN: 978-1-4939-3622-9

  • eBook Packages: Springer Protocols

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