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Fig.1 Western blot analysis TritonX-100 insoluble cytoskeletal fraction (lane 1) and concentrated conditioned medium (lane 2) prepared from DJM-1 cells were immunoblotted with the C34 antibody (1:200 dilution). The C34 antibody detected a band at approximately 180 kDa in lane 1. This antibody also reacted with a 120-kDa shed ectodomain of BP180 in lane 2. Polypeptiedes were separated by SDS-PAGE (7.5% separating gel).
Fig.3 Immunofluorescence microscopy of human skin A human skin section was stained with C34 antibody at 1:200 dilution. The antibody revealed the location of BP180 molecules at the dermal-epidermal junction (arrow). E: epidermis, D: dermis. Bar = 50 um. Frozen sections were prepared as described previously (ref. 3).
Fig.2 Location of the epitope for the C34 antibody The C34 antibody does not react with the 100-kDa extracellular fragment of BP180, which lacks the COOH-terminal portion (ref. 1). The result indicates that the C34 antibody recognizes an epitope locates at the COOH-terminal portion of about 20 kDa. ICD, TM and ECD represent for intracellular, transmembrane and extracellular domains. Collagenous domains are shown by gray boxes.
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