Desmin, human recombinant

Molecular Weight 53,539.6 (calculated from sequence)55,000 (determined by SDS gelelectrophoresis) Application Protein standard in 1D and 2D SDS gelelectrophoresis, Immunoassays, Immunization Isoelectric Point pI 5.16 (calculated from sequence) Reconstitution Reconstitute with 70 μl distilled water (final volume 100 μl)Reconstitute with 175 μl distilled water (final volume 250 μl)Final solution: 30 mM Tris/HCI pH 8, 9.5 M urea, 2 mM DTT, 2 mM EDTA, 10 mM methylammonium chloride; protein concentration: 1 mg/ml Storage At 2-8°C (lyoph.); at -20°C (reconstituted) Reconstitution to filaments: after desmin is dissolved in 9.5 M urea buffer (see above), protofilaments and filament complexes are obtained by dialyzing the resulting polypeptide solution stepwise to a concentration of 4M urea and then to low salt condition (50 mM NaCl, 2 mM dithiothreitol, 10 mM Tris-HCl, pH 7.4). For immunization purposes, the solution can be further dialyzed against PBS (phosphate buffered saline, e.g. Dulbecco's PBS). Hatzfeld M. and Franke W.W. (1985). J. Cell Biol. 101, 1826-1841 Hatzfeld M. et al. (1987). J. Mol. Biol. 197, 237-255