Molecular Weight 53,000
Application Protein standard in 1D and 2D SDS gelelectrophoresis, Immunoassays, Immunization
Isoelectric Point pI 5.4
Storage At 2-8°C (lyoph.); at -20°C (reconstituted)
ReconstitutionReconstitute with 80 μl distilled water (final volume 100 μl)Reconstitute with 200 μl distilled water (final volume 250 μl)Final solution: 10 mM sodium phosphate buffer pH 7.5, 6M urea, 2 mM DTT, 1 mM EDTA, 10 mM methylammonium chloride; protein concentration: 1 mg/ml (determined by Bradford method)
Reconstitution to filaments is performed by dissolving in 6 M urea buffer (see above) at concentrations of approx. 0.5 mg/ml. Protofilaments and filament complexes are obtained by dialyzing the resulting polypeptide solution stepwise to a concentration of 4 M 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 WW (1985). J Cell Biol 101, 1826-1841
Hatzfeld M et al. (1987). J Mol Biol 197, 237-255