Localization of HBP by immunofluorescence microscopy in HL60 cells.

Indirect immunolocalization of HBP and markers for azurophilic granules and secretory vesicles. HL60 cells were fixed, permeabilized, and stained, as described in “Materials and methods,” before attachment to poly-L-lysine–coated coverslips and recording of images. (A-C) Cells were incubated with a rabbit anti-HBP antibody and a mouse monoclonal antibody against the azurophilic granule marker, CD63. (D-F) Cells were incubated with a rabbit anti-HBP antibody and a mouse monoclonal antibody against the secretory vesicle marker, CD35. Thereafter, the cells were stained with FITC-labeled anti-rabbit and Cy3-labeled anti-mouse secondary antibodies. Staining of HBP (A, D), CD63 (B), and CD35 (E). Corresponding Nomarski images (C, F). Results are representative of 3 separate experiments. Bar = 10 μM.