Immunological characterization of exocyst complex subunits in cell differentiation.
We have generated monoclonal antibodies (MAbs) against three proteins sec6, sec15, and exo84. These proteins have been shown to be components of the exocyst complex, a macromolecule required for many biological processes such as kidney epithelial formation and neuronal development. These antibodies can detect the three proteins by enzyme-linked immunoadsorbent assay (ELISA), Western blotting, immunofluorescence microscopy, and immunoprecipitation. Using these antibodies, we found that the three proteins have similar subcellular localization which changes upon cell differentiation. These three proteins also co-immunoprecipitate with each other. These results suggest that at least three exocyst subunits associate with each other in vivo and redistribute in response to cell differentiation. In the future, these antibodies should be useful in the cell biological and functional analysis of the exocyst complex under physiological and pathological conditions.
Wang
Sheng
S
Department of Cell Biology and Neurosciences, Rutgers University, Piscataway, NJ 08854, USA.
Hsu
Shu C
SC
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism; D002352:Carrier Proteins / Q000737:chemistry*; D002454:Cell Differentiation; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D007668:Kidney / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry*; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D009474:Neurons / Q000378:metabolism; D011233:Precipitin Tests; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry; D033921:Vesicular Transport Proteins
D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D018076:DNA, Complementary; C490685:EXOC3 protein, human; C480857:EXOC8 protein, human; C096102:Exoc3 protein, rat; C109899:Exoc6 protein, rat; D008565:Membrane Proteins; D011994:Recombinant Proteins; C490683:Sec6l1 protein, mouse; D033921:Vesicular Transport Proteins