Generation of monoclonal antibodies specific for desmoglein family members.
Desmosomes are the most prominent cell-cell junctions in most epithelial cells and serve to link the intermediate filament cytoskeletons of adjacent cells. Desmogleins and desmocollins are the transmembrane core of the desmosome and both are members of the cadherin family of cell-cell adhesion molecules. In the skin, the three desmoglein gene products (Dsg 1, 2, and 3) are expressed in a stratification dependent manner, and therefore contribute to compositionally different desmosomes throughout the differentiating layers. In this study we generated a panel of monoclonal antibodies (MAbs) specific for each of the desmoglein gene products and evaluated their usefulness in a number of immunological procedures including immunoblotting, immunoprecipitation, and immunofluorescence. In addition, we showed that these antibodies are useful for immunoprecipitating desmogleins from cell extracts prepared in 0.1% Empigen BB, a zwitterionic detergent capable of solubilizing the desmosomal structure. Identification of conditions that solubilize the desmosome and allow the use of immunological reagents will help facilitate an increased understanding of desmosome assembly and regulation.
Wahl
James K
JK
Department of Oral Biology and Eppley Cancer Center, College of Dentistry, University of Nebraska Medical Center, 986805 Nebraska Medical Center, Omaha, NE 68198, USA. jwahl@unmc.edu
D000911:Antibodies, Monoclonal / Q000378:metabolism*; D015820:Cadherins / Q000378:metabolism; D002448:Cell Adhesion; D002454:Cell Differentiation; D003598:Cytoskeletal Proteins / Q000737:chemistry* / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D051187:Desmocollins; D051183:Desmoglein 1; D051184:Desmoglein 3; D051182:Desmogleins; D051180:Desmoplakins; D003902:Detergents / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004847:Epithelial Cells / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D008856:Microscopy, Fluorescence; D011233:Precipitin Tests; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D015820:Cadherins; D003598:Cytoskeletal Proteins; D018076:DNA, Complementary; C071894:DSG3 protein, human; D051187:Desmocollins; D051183:Desmoglein 1; D051184:Desmoglein 3; D051182:Desmogleins; D051180:Desmoplakins; D003902:Detergents; D011994:Recombinant Proteins