Staining of the Midbody by an Anti-digoxin-specific Antibody

Abstract

SUMMARY Using RNA in situ hybridization to reveal cytoplasmic localization patterns of mRNAs in cultured cells, we noted unexpected staining of a cytoplasmic component in telophase cells. Control experiments revealed that the anti-digoxin-specific antibody was responsible for this staining. Because the staining was observed only at a position where both daughter cells are still connected, we identified the stained component as the midbody. This was confirmed by double staining of cells with anti-digoxin and anti-␣ -tubulin antibodies. We concluded that anti-digoxin-specific antibody shows crossreactivity with a component present in the midbody.

Methods for visualizing RNA processing and transport pathways in living cells

“…Fixation and cell pretreatment procedures may disturb the localization of RNAs and to some extent deteriorate cell morphology. Furthermore, some of the antibodies that are routinely used to detect haptenized probes following hybridization are shown to (cross)react with endogenous cytoplasmic components (Chevalier et al 1997;Dirks and Raap 1998). In addition, information about the dynamics of RNA synthesis and transport can be obtained best when these processes are studied in living cells.…”