ProteinImmunolocalizationinMaizeTissues

The  analysis of gene expression at transcript and at protein level is of  outstanding importance in plant developmental biology. Proteins can be  localized with subcellular resolution by immunolocalization using  specific antibodies or generating reporter lines carrying the specific  protein fused to a fluorescent protein. Immunolocalization is  particularly suitable to confirm the expression pattern of transgenic  reporter lines. It also represents a valid alternative, especially for  plants such as maize, for which transformation is time consuming and  still often unsuccessful, by-passing also some side-effects of fusion  proteins. Indeed, the availability of specific antibodies for  immunolocalizations and observations of maize tissues under a confocal  microscope is a powerful tool for studying protein targeting to  different cellular compartments.  

In the following chapter we describe the complete procedure for the  localization of proteins in different maize tissues both at cellular and  sub-cellular level, using polyclonal or monoclonal antibodies. Tissues  can be embedded in different substrates, such as paraplast, PEG400 and  agarose, depending on the tissue and the desired use: epifluorescence or  confocal microscope observations. An additional protocol for the  analysis of the nuclear distribution of modified histones in squashed  maize root apexes is also presented.