The Journal of Cell Biology, Vol 92, 92-107, Copyright © 1982 by The Rockefeller University Press

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Antibodies to the Golgi complex and the rough endoplasmic reticulum

D Louvard, H Reggio and G Warren

Rabbits were immunized with membrane fractions from either the Golgi complex or the rough endoplasmic reticulum (RER) by injection into the popliteal lymph nodes. The antisera were then tested by indirect immunofluorescence on tissue culture cells or frozen, thin sections of tissue. There were may unwanted antibodies to cell components other than the RER or the Golgi complex, and these were removed by suitable absorption steps. These steps were carried out until the pattern of fluorescent labeling was that expected for the Golgi complex or RER. Electron microscopic studies, using immunoperoxidase labeling of normal rat kidney (NRK) cells, showed that the anti-Golgi antibodies labeled the stacks of flattened cisternae that comprise the central feature of the Golgi complex, many of the smooth vesicles around the stacks, and a few coated vesicles. These antibodies were directed, almost entirely, against a single polypeptide with an apparent molecular weight of 135,000. The endoplasmic reticulum (ER) in NRK cells is an extensive, reticular network that pervades the entire cell cytoplasm and includes the nuclear membrane. The anit-RER antibodies labeled this structure alone at the light and electron microscopic levels. They were largely directed against four polypeptides with apparent molecular weights of 29,000, 58,000, 66,000, and 91,000. Some examples are presented, using immunofluorescence microscopy, where these antibodies have been used to study the Golgi complex and RER under a variety of physiological and experimental condition . For biochemical studies, these antibodies should prove useful in identifying the origin of isolated membranes, particularly those from organelles such as the Golgi complex, which tend to lose their characteristic morphology during isolation.
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• Wong, S. H., Xu, Y., Zhang, T., Griffiths, G., Lowe, S. L., Subramaniam, V. N., Seow, K. T., Hong, W. (1999). GS32, a Novel Golgi SNARE of 32 kDa, Interacts Preferentially with Syntaxin 6. Mol. Biol. Cell 10: 119-134 [Abstract] [Full Text]  
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• Xu, Y., Wong, S. H., Tang, B. L., Subramaniam, V. N., Zhang, T., Hong, W. (1998). A 29-Kilodalton Golgi Soluble N-Ethylmaleimide-sensitive Factor Attachment Protein Receptor (Vti1-rp2) Implicated in Protein Trafficking in the Secretory Pathway. J. Biol. Chem. 273: 21783-21789 [Abstract] [Full Text]  
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• Rouiller, I., Brookes, S. M., Hyatt, A. D., Windsor, M., Wileman, T. (1998). African Swine Fever Virus Is Wrapped by the Endoplasmic Reticulum. J. Virol. 72: 2373-2387 [Abstract] [Full Text]  
• Charrasse, S, Schroeder, M, Gauthier-Rouviere, C, Ango, F, Cassimeris, L, Gard, D., Larroque, C (1998). The TOGp protein is a new human microtubule-associated protein homologous to the Xenopus XMAP215. J. Cell Sci. 111: 1371-1383 [Abstract]  
• Levi, S, Vannier, C, Triller, A (1998). Strychnine-sensitive stabilization of postsynaptic glycine receptor clusters. J. Cell Sci. 111: 335-345 [Abstract]  
• Rottger, S, White, J, Wandall, H., Olivo, J., Stark, A, Bennett, E., Whitehouse, C, Berger, E., Clausen, H, Nilsson, T (1998). Localization of three human polypeptide GalNAc-transferases in HeLa cells suggests initiation of O-linked glycosylation throughout the Golgi apparatus. J. Cell Sci. 111: 45-60 [Abstract]  
• Lander, C., Zhang, H., Hockfield, S. (1998). Neurons Produce a Neuronal Cell Surface-Associated Chondroitin Sulfate Proteoglycan. J. Neurosci. 18: 174-183 [Abstract] [Full Text]  
• Gonatas, J., Chen, Y., Stieber, A, Mourelatos, Z, Gonatas, N. (1998). Truncations of the C-terminal cytoplasmic domain of MG160, a medial Golgi sialoglycoprotein, result in its partial transport to the plasma membrane and filopodia. J. Cell Sci. 111: 249-260 [Abstract]  
• Shenkman, M., Ayalon, M., Lederkremer, G. Z. (1997). Endoplasmic reticulum quality control of asialoglycoprotein receptor H2a involves a determinant for retention and not retrieval. Proc. Natl. Acad. Sci. USA 94: 11363-11368 [Abstract] [Full Text]  
• Xu, Y., Wong, S. H., Zhang, T., Subramaniam, V. N., Hong, W. (1997). GS15, a 15-Kilodalton Golgi Soluble N-Ethylmaleimide-sensitive Factor Attachment Protein Receptor (SNARE) Homologous to rbet1. J. Biol. Chem. 272: 20162-20166 [Abstract] [Full Text]  
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• Lowe, S., Wong, S., Hong, W (1996). The mammalian ARF-like protein 1 (Arl1) is associated with the Golgi complex. J. Cell Sci. 109: 209-220 [Abstract]  
• Hoe, M. H., Slusarewicz, P., Misteli, T., Watson, R., Warren, G. (1995). Evidence for Recycling of the Resident medial/trans Golgi Enzyme, N-Acetylglucosaminyltransferase I, in ldlD Cells. J. Biol. Chem. 270: 25057-25063 [Abstract] [Full Text]  
• Goodnight, J., Mischak, H., Kolch, W., Mushinski, J. F. (1995). Immunocytochemical Localization of Eight Protein Kinase C Isozymes Overexpressed in NIH 3T3 Fibroblasts. J. Biol. Chem. 270: 9991-10001 [Abstract] [Full Text]  
• Subramaniam, V., Krijnse-Locker, J, Tang, B., Ericsson, M, Yusoff, A., Griffiths, G, Hong, W (1995). Monoclonal antibody HFD9 identifies a novel 28 kDa integral membrane protein on the cis-Golgi. J. Cell Sci. 108: 2405-2414 [Abstract]  
• Rabouille, C, Hui, N, Hunte, F, Kieckbusch, R, Berger, E., Warren, G, Nilsson, T (1995). Mapping the distribution of Golgi enzymes involved in the construction of complex oligosaccharides. J. Cell Sci. 108: 1617-1627 [Abstract]  
• Horvat, R, Palade, G. (1995). The functional thrombin receptor is associated with the plasmalemma and a large endosomal network in cultured human umbilical vein endothelial cells. J. Cell Sci. 108: 1155-1164 [Abstract]  
• Gonatas, J., Mourelatos, Z, Stieber, A, Lane, W., Brosius, J, Gonatas, N. (1995). MG-160, a membrane sialoglycoprotein of the medial cisternae of the rat Golgi apparatus, binds basic fibroblast growth factor and exhibits a high level of sequence identity to a chicken fibroblast growth factor receptor. J. Cell Sci. 108: 457-467 [Abstract]  
• Pryde, J. (1994). A group of integral membrane proteins of the rat liver Golgi contains a conserved protein of 100 kDa. J. Cell Sci. 107: 3425-3436 [Abstract]  
• Lecureux, L., Wattenberg, B. (1994). The regulated degradation of a 3-hydroxy-3-methylglutaryl-coenzyme A reductase reporter construct occurs in the endoplasmic reticulum. J. Cell Sci. 107: 2635-2642 [Abstract]  
• Mollat, P, Fournier, A, Yang, C., Alsat, E, Zhang, Y, Evain-Brion, D, Grassi, J, Thang, M. (1994). Species specificity and organ, cellular and subcellular localization of the 100 kDa Ras GTPase activating protein. J. Cell Sci. 107: 427-435 [Abstract]  
• Johnston, P., Stieber, A, Gonatas, N. (1994). A hypothesis on the traffic of MG160, a medial Golgi sialoglycoprotein, from the trans-Golgi network to the Golgi cisternae. J. Cell Sci. 107: 529-537 [Abstract]  
• Roa, M, Cornet, V, Yang, C., Goud, B (1993). The small GTP-binding protein rab6p is redistributed in the cytosol by brefeldin A. J. Cell Sci. 106: 789-802 [Abstract]  
• Pieters, J, Bakke, O, Dobberstein, B (1993). The MHC class II-associated invariant chain contains two endosomal targeting signals within its cytoplasmic tail. J. Cell Sci. 106: 831-846 [Abstract]  
• Schweizer, A, Ericsson, M, Bachi, T, Griffiths, G, Hauri, H. (1993). Characterization of a novel 63 kDa membrane protein. Implications for the organization of the ER-to-Golgi pathway. J. Cell Sci. 104: 671-683 [Abstract]  
• Heusser, S., Colin, S., Figiel, A., Huet, C., Keller, J. M., Pornet, P., Robine, S., Vandamme, J., Vandekerckhove, J., Dauca, M. (1992). Amphibian intestinal villin: isolation and expression during embryonic and larval development. J. Cell Sci. 103: 699-708 [Abstract]  
• Maunoury, R, Robine, S, Pringault, E, Leonard, N, Gaillard, J., Louvard, D (1992). Developmental regulation of villin gene expression in the epithelial cell lineages of mouse digestive and urogenital tracts. Development 115: 717-728 [Abstract]  
• Miller, S C, Pavlath, G K, Blakely, B T, Blau, H M (1988). Muscle cell components dictate hepatocyte gene expression and the distribution of the Golgi apparatus in heterokaryons.. Genes Dev. 2: 330-340 [Abstract]  
• Lipsky, N., Pagano, R. (1985). A vital stain for the Golgi apparatus. Science 228: 745-747 [Abstract]  
• Carreno, S., Gouze, M.-E., Schaak, S., Emorine, L. J., Maridonneau-Parini, I. (2000). Lack of Palmitoylation Redirects p59Hck from the Plasma Membrane to p61Hck-positive Lysosomes. J. Biol. Chem. 275: 36223-36229 [Abstract] [Full Text]  

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