Vimentin, human recombinant, 250 µg
- Recombinant human vimentin
- Protein standard
|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||57 kDa|
|Isoeletric point||pI 5.3|
|Purity||> 95% (determined by SDS gelelectrophoresis)|
|Reconstitution||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
|Application||Protein standard in 1D and 2D SDS gelelectrophoresis, immunoassays and immunization|
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
Q & A's
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- Reconsitute the lyophilized material in distilled water to a protein concentration of 1 mg/ml and a urea concentration of 9.5 M.
- Dialyze against 6 M urea buffer (down to 6 M urea vimentin is still present in the monomeric form) for at least 2 h at RT.
- Dialyze against 4M urea buffer (at 4 M urea dimers and protofilaments are formed) for at least 4 h at RT or overnight at 4°C.
- Dialyze to low urea/low salt buffer (at least 2 buffer changes to obtain intermediate filaments) containing 50 mM NaCl, 2 mM dithiothreitol, 10 mM Tris-HCl, pH 7.4, or filament forming buffer (suggested e.g. by Herrmann et al. 1996, J Mol Biol 264, 933-953) containing 25 mM Tris-HCl pH 7.5, 0.1 mM dithiothreitol, 160 mM NaCl- for immunization purposes dialyze to e.g. PBS.
- Dialysis in steps against large buffer volumes is important in order to obtain filament complexes and avoid formation of amorphous aggregates.
- Purified, lyophilized
- Mouse monoclonal
- Suitable for ICC/IF, IHC and WB
- Reacts with amphibia, bovine, chicken, human, mouse and monkey
- Isotype: IgG2a