Desmin, chicken gizzard, 250 µg

Cat. No. 62005

Available, delivery time 1-7 days


Key Features
  • Purified chicken gizzard desmin
  • Protein standard
Product description
Quantity 250 µg
Storage Lyophilized at 2-8°C; reconstituted at -20°C (avoid freeze/thaw cycles)
Intended use Research use only
Source Chicken gizzard
Molecular weight 53 kDa
Isoeletric point pI 5.4
Purity > 98% (determined by SDS gelelectrophoresis)
Reconstitute with 200 µl distilled water (final volume 250 µl).
Final solution: 10 mM sodium phosphate buffer pH 7.5, 6 M urea, 2 mM DTT, 1 mM EDTA, 10 mM methylammonium chloride;
Protein concentration: 1 mg/ml (determined by Bradford method).
Application Protein standard in 1D and 2D SDS gelelectrophoresis, immunoassays and immunization
Protein standard for immunoblotting, immunization and immunoassays.
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
Q & A's

Please enter a valid email address.
Please enter a valid question.
Fields marked with * are mandatory.

There aren't any asked questions yet.
Customer Reviews

0 of 0 reviews

Leave a review!

Share your experiences with other customers.

Our chicken Desmin was purified according to the following protocol:
  • Storage of chicken gizzard in liquid Nitrogen at -196°C (-320 °F).
  • Homogenization in salt solution.
  • Solubilization in 6-9 M urea solution.
  • Ion exchange chromatography in 6-9 M urea solution for > 12 h.
  • Elution with increasing salt concentration.
  • HPLC purification in 0.1% trifluoroacetic acid and acetonitril at pH 2.0.
  • Vacuum rotation and dialysis.
  • Filtration through 0.45 µm filter.
  • Lyophilization.

Our chicken Desmin was purified using strong denaturing conditions and HPLC:
Raw material: frozen chicken gizzards obtained from the local abattoir, Mannheim / Germany;
  • Homogenization in buffer I (50 mM Imidazol, pH 7.4;  2 mM DTT, 50 µM Pefabloc).
  • Preparation of cytoskeletal material (several wash and centrifugation steps in buffer I).
  • Extraction of cytoskeletal material with urea (buffer I plus 8 M urea).
  • Chromatographic separation of urea extract on Sepharose S in presence of urea (elution of desmin containing fractions with NaCl-gradient: 100-500 mM NaCl), concentrating and dialysis of desmin-containing fractions.
  • Re-chromatography of desmin-containing fractions on DEAE cellulose, elution with 50 mM NaCl.
  • Concentrating desmin-containing fractions by precipitation with 100% ethanol.
  • Chromatography by HPLC (reverse phase: TFA/acetonitril).
  • Evaporation of dissolving solution.
  • Analysis by SDS-PAGE and Western blotting.
  • Determination of protein concentration by Bradford method.
  • Solubilization in urea buffer and lyophilization.