Antidesmin mouse monoclonal antibody clone d33

HSMM and SkMc myocytes were cultured for 5–7 days in differentiation medium on 4-well Lab-Tek II chamber slides (Electron Microscopy Sciences, Hatfield, Pennsylvania, USA). Next, the cells were fixed with ice-cold 100% methanol for 30 min at –20°C and stained for 1 h with antidesmin mouse monoclonal antibody (clone D33, Dako, Denmark, 1:25 dilution in 5% normal donkey serum in PBS) followed by a 1 h incubation with Alexa Fluor488 donkey antimouse IgG (Invitrogen, 1:50 dilution in 5% normal donkey serum in PBS). Next, the cell slides were covered with Vectashield mounting medium with 4',6' diamino-2-phenylindole·2HCl (DAPI) (Vector Laboratories, Burlingame, California, USA) and evaluated under Nikon Eclipse TE2000 fluorescent microscope.

Anti-podoplanin rabbit polyclonal antibody (KS-17) was from Sigma (St. Louis, MO). Anti-multi-cytokeratin mouse monoclonal antibody (RTU-AE1/AE3) was from Novocastra (Newcastle, UK). Anti-desmin mouse monoclonal antibody (clone D33) was from DAKO (Carpinteria, CA). Anti-calretinin rabbit monoclonal antibody (clone SP13) was from Abcam (Cambridge, MA). Anti-S-100 polyclonal antibody was from DAKO. Anti-C-ERC/mesothelin monoclonal antibody was from IBL (Takasaki, Gunma, Japan). Anti-WT1 rabbit polyclonal antibody (C-19) was from Santa Cruz (Santa Cruz, CA). Secondary antibody was Histofine Simplestain rat MAX-PO (Multi) from Nichirei Bioscience (Tokyo).