Vimentin, human recombinant, 100 µg

Vimentin, human recombinant, 100 µg View more

100 µg

250 µg

Cat. No: 62215

Prices excl. tax

Available, delivery time 1-7 days

$364.00*

Datasheet

Key Features

Recombinant human vimentin
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	57 kDa
Isoeletric point	pI 5.3
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

Background

Protein standard for immunoblotting, immunization and immunoassays.

Reconstitution to filaments: after vimentin 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 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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Datasheet

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FAQs

Our human recombinant vimentin was lyophilized in urea buffer. Vimentin other intermediate filament proteins are insoluble under physiological buffer conditions (like PBS) or low salt conditions. This is a prominent feature of this protein family. There is some solubility in Tris-buffer, however, at very low protein concentration.

To obtain filaments from Cat. No. 62015 we suggest the following procedure:

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.

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