Nickel-Intensified ABC-HRP Antibody Staining of Imaginal Discs in Drosophila
This protocol was adapted from “Imaginal Discs,” Chapter 10, in Drosophila Protocols (eds. Sullivan et al.). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2000.INTRODUCTION
This protocol describes a sensitive method for labeling Drosophila imaginal disc tissues with a single antiserum. Labeling is visualized using a nickel-intensified avidin/biotinylated enzyme complex-horseradish peroxidase (ABC-HRP) staining procedure. This method is more sensitive, and has a lower signal-to-noise ratio, than fluorescent labeling. Although these methods are useful for visualizing genetic mosaic clones in wandering larval stage imaginal discs and pupal tissues from Drosophila, the immunohistological techniques are generally applicable to the description of protein expression.
