Journal: The Journal of Cell Biology

Article Title: Insoluble γ-Tubulin–Containing Structures Are Anchored to the Apical Network of Intermediate Filaments in Polarized Caco-2 Epithelial Cells

doi:

Figure Lengend Snippet: Colocalization of γ-tubulin and CK19 IF at EM level. CACO-2 cell monolayers were grown on filters as described above and fixed in methanol. CK19 was localized with Nanogold™ gold particles followed by silver developing, while γ-tubulin was localized using a standard immunoperoxidase procedure. (a) CK19-Nanogold™ signal was developed with silver for 20 min to attain 200–400 nm particles visible at low magnification. (b) The CK19-Nanogold™ signal was developed with silver for only 5 min, so that the particles were in the range 15–20 nm, and colocalized with γ-tubulin detected by immunoperoxidase (diffuse stain) reaction confined within a matrix of cross-linked albumin. Note that the minimum distance from gold particles to peroxidase stain is 10–20 nm (arrows). Bar: (a) 1.6 μm; (b) 0.1 μm.

Article Snippet: Low molecular mass Fab goat anti–rabbit IgG coupled to peroxidase was purchased from Protos Immunoresearch.

Techniques: Staining