Immunolocalization of Arabinogalactan Proteins and Pectins in Plant Tissue

Take a slide with reaction wells containing thin sections of different plant tissues. The tissues are fixed and embedded inside a resin for immunostaining.

Place the slide inside a dark and humid incubation chamber to prevent light exposure and reagent evaporation during subsequent steps.

Introduce a blocking solution to prevent non-specific antibody interaction.

Wash to remove excess blocking solution, then add antigen-specific primary antibodies and incubate.

The antibodies bind to their target antigens, namely arabinogalactan proteins and pectins in the cell wall.

Introduce fluorophore-conjugated secondary antibodies, which bind to the primary antibodies.

Wash to remove the unbound antibodies, and apply a fluorescent dye that binds to cellulose, labeling the cell wall.

Add a mounting medium and seal with a coverslip.

Under a microscope, fluorescence from the dye and the bound antibodies aid in visualizing cellulose in plant cell walls colocalized with arabinogalactan proteins or pectin, exhibiting their tissue-specific distribution.