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Keratin K18, human recombinant, 100 µg

Key Features
  • Recombinant human keratin K18 (formerly also designated cytokeratin 18)
  • Protein standard
Product description
Quantity 100 µg
Storage Lyophilized at 2-8°C; reconstituted at -20°C (avoid freeze/thaw cycles)
Intended use Research use only
Source Human recombinant, produced in E. coli
Molecular weight 45 kDa
Isoeletric point pI 5.7
Purity > 95% (determined by SDS gelelectrophoresis)
Reconstitution Reconstitute with 70 µl distilled water (final volume 100 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
Application Protein standard in 1D and 2D SDS gelelectrophoresis, immunoassays and immunization
Synonym Cytokeratin 18
Background
Human recombinant Keratin K18 for use in immunoblotting and ELISA.
Reconstitution to filaments is performed by mixing equimolar amounts of keratins of type I and type II at concentrations of approx. 0.5 mg/ml, both 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 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 W.W. (1985). J. Cell Biol. 101, 1826-1841
- Hatzfeld M. et al. (1987). J. Mol. Biol. 197, 237-255
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FAQs
Reconstitution to filaments is performed by mixing equimolar amounts of cytokeratins of type I and type II at concentrations of approx. 0.2-0.5 mg/ml, both dissolved in 9.5 M urea buffer: e.g. cytokeratin 18 solution in urea buffer plus cytokeratin 8 solution in urea buffer (both with the same protein concentration!) Protofilaments and filament complexes are obtained by dialyzing the resulting polypeptide solution stepwise to a concentration of 4 M urea buffer  (for approx. 4 h at RT) and then to low salt condition (overnight at 2-8°C, using e.g. 50 mM NaCl, 2 mM dithiothreitol, 10 mM Tris-HCl, pH 7.4). Use a large excess of dialyzing buffer (e.g. x500). For immunization purposes, the solution can be further dialyzed against PBS (phosphate buffered saline, e.g. Dulbecco's PBS). For storage of the dialyzed solution we recommend division in suitable aliquots and freezing below -50°C. Avoid repeated freezing / thawing cycles.

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