Labeling Microtubules (Molecular Dynamics Inc. )
Microtubules are involved in many aspects of cell motion including propulsion, mitosis, growth, and organelle transport. They are composed of a- and ß-tubulin, and are about 25 nm in outer diameter. Microtubules are easily stained using an anti-tubulin primary antibody and a corresponding secondary antibody conjugated to any fluorochrome. Their regular size and fine structure provide beautiful images. This procedure reveals smooth and linear microtubules and can be used with a wide usr/localiety of other antigens as well.
Analysis of Intracellular Organelles by Flow Cytometry or Microscopy (CPC Protocols)
Functional analysis of cellular organelles can be accomplished by staining cells with suitable organelle-specific dyes and then analyzing the fluorescence of the stained cells with a flow cytometer. With this methodology it is possible to resolve suspected heterogeneity in organelle function or content within a population of cells. Flow cytometry does not provide morphological information; if that is desired, quantitative microscopyusing a video microscope with digital image analysis system, or a confocal microscopeshould be employed
Preparation of tubulin (Salmon Lab)
Simple procedure with high yielding
Preparation of Rat Liver Cell Cytosol and/or Organelle Fractions (Salmon Lab)
Adrenal Chromaffin Granule (chromaffin vesicle) Preparation (Daniel T. O'Connor)
This method is usually used to prepare granules from freshly obtained (never frozen!) bovine adrenal glands, but can also be used to prepare chromaffin granules from fresh human pheochromocytoma.
Lysosome Isolation in Isotonic Sucrose (William H. Heidcamp)
Em Observations of Microsomes (William H. Heidcamp)
Liposome Isolation from Bacteria (Hancock Lab) (Accessible only by IE)
