Abstract
Antibodies are powerful tools for studying protein localization and function. Here we describe methods for two of the more challenging applications: immunolocalization and chromatin immunoprecipitation (ChIP). Immunolocalization can be carried out on nearly any maize tissue, but we focus here on localizing proteins to male meiocytes, which are large cells with relatively porous cell walls that make them ideal for this purpose. ChIP is a biochemical technique for identifying DNA sequences that interact with chromatin proteins such as histones. We describe a thoroughly tested method for native ChIP (useful for histones) and one method for crosslinking ChIP, which can be used for proteins that bind to the surface of DNA.
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Abbreviations
- ChIP:
-
Chromatin immunoprecipitation
- LN2 :
-
Liquid nitrogen
- ME:
-
2-Mercaptoethanol
- nChIP:
-
Native ChIP
- PAS:
-
Protein A sepharose
- PBS:
-
Phosphate-buffered saline
- PI:
-
Protease inhibitor tablet
- PMSF:
-
Phenylmethanesulphonylfluoride
- SDS:
-
Sodium dodecyl sulfate
- xChIP:
-
Crosslinking ChIP
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Li, X., Topp, C.N., Dawe, R.K. (2012). Maize Antibody Procedures: Immunolocalization and Chromatin Immunoprecipitation. In: Bass, H., Birchler, J. (eds) Plant Cytogenetics. Plant Genetics and Genomics: Crops and Models, vol 4. Springer, New York, NY. https://doi.org/10.1007/978-0-387-70869-0_11
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DOI: https://doi.org/10.1007/978-0-387-70869-0_11
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