Encyclopedia of Experiments: Immunology
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Transcript
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.
