Tagging an organelle
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《细胞学杂志》
.To the "raw" antibody solution they added debris they had initially separated from the Golgi membranes. This junk was mainly plasma membranes and bits of endoplasmic reticulum. After letting the combination incubate, they again removed the membrane gunk—and in the process eliminated many of the unwanted antibodies (Louvard et al., 1982). The researchers then performed the step again with rat plasma, which sopped up antibodies against secretory proteins. Using immunofluorescence, the researchers showed that the leftover mixture labeled only the perinuclear region, where the Golgi complex forms. Meanwhile, cells tagged with the "raw" antibody concoction glowed all over. "This was a dramatic demonstration that you could make high-affinity antibodies to organelles," says Warren. Cell biologists expressed their approval in the usual way, he says: "They asked us for samples."
Raw anti-Golgi antibodies (top) are more specific after extraneous antibodies are removed (bottom).
LOUVARD
The researchers determined that the antibodies were recognizing one Golgi protein—though they weren't sure of its identity or location. Subsequent work revealed that it was mannosidase II, a key Golgi enzyme. Louvard and colleagues applied the same technique to uncover four markers for the endoplasmic reticulum. Their discovery helped researchers better understand the anatomy and activity of the Golgi complex. But it also sparked a controversy—over whether the complex forms spontaneously or requires a template—that hasn't abated today (Wells, 2001).
Bader, D., et al. 1982. J. Cell Biol. 95:763–770.
de Camilli, P., et al. 1983a. J. Cell Biol. 96:1337–1354.
de Camilli, P., et al. 1983b. J. Cell Biol. 96:1355–1373.
Huttner, W.B., et al. 1983. J. Cell Biol. 96:1374–1388.
Levine, J., and M. Willard. 1981. J. Cell Biol. 90:631–643.
Louvard, D., et al. 1982. J. Cell Biol. 92:92–107.
Schnitzer, J., et al. 1981. J. Cell Biol. 90:435–447.
Skene, J.H.P., and M. Willard. 1981. J. Cell Biol. 89:96–103.
Weiss, R.A., et al. 1984. J. Cell Biol. 98:1397–1406.
Wells, W.A. 2001. J. Cell Biol. 155:498–499.
Woodcock-Mitchell, J., et al. 1982. J. Cell Biol. 95:580–589.
Yen, S.-H., and K.L. Fields. 1981. J. Cell Biol. 88:115–126.(In the early 1980s, cell biologists kept)
Raw anti-Golgi antibodies (top) are more specific after extraneous antibodies are removed (bottom).
LOUVARD
The researchers determined that the antibodies were recognizing one Golgi protein—though they weren't sure of its identity or location. Subsequent work revealed that it was mannosidase II, a key Golgi enzyme. Louvard and colleagues applied the same technique to uncover four markers for the endoplasmic reticulum. Their discovery helped researchers better understand the anatomy and activity of the Golgi complex. But it also sparked a controversy—over whether the complex forms spontaneously or requires a template—that hasn't abated today (Wells, 2001).
Bader, D., et al. 1982. J. Cell Biol. 95:763–770.
de Camilli, P., et al. 1983a. J. Cell Biol. 96:1337–1354.
de Camilli, P., et al. 1983b. J. Cell Biol. 96:1355–1373.
Huttner, W.B., et al. 1983. J. Cell Biol. 96:1374–1388.
Levine, J., and M. Willard. 1981. J. Cell Biol. 90:631–643.
Louvard, D., et al. 1982. J. Cell Biol. 92:92–107.
Schnitzer, J., et al. 1981. J. Cell Biol. 90:435–447.
Skene, J.H.P., and M. Willard. 1981. J. Cell Biol. 89:96–103.
Weiss, R.A., et al. 1984. J. Cell Biol. 98:1397–1406.
Wells, W.A. 2001. J. Cell Biol. 155:498–499.
Woodcock-Mitchell, J., et al. 1982. J. Cell Biol. 95:580–589.
Yen, S.-H., and K.L. Fields. 1981. J. Cell Biol. 88:115–126.(In the early 1980s, cell biologists kept)