Vimentin-Chromobody plasmid

Description
DNA plasmid encoding for anti-Vimentin VHH (anti-Vimentin Nanobody) fused to TagGFP2.

Specificity
Vimentin intermediate filament protein, tested in human, hamster & dog cells

Applications

Trace endogenous vimentin in real-time
Live cell imaging
Monitor epithelial-mesenchymal transition in live cells

Vimentin-Chromobody plasmid
Vimentin-Chromobody plasmid (TagGFP)

Product Size Code Price Buy
Product Vimentin-Chromobody® plasmid (TagGFP2) Size 20 µg Code vcg Price $ 999
Buy +

Vimentin-Chromobody format

 

Vimentin-Chromobody
plasmid (TagGFP2)

Vector type

mammalian
expression vector

Promoter

constitutive CMV IE

Reporter

TagGFP2

Codon usage

mammalian

Selection

Kan/Neo

Nuclear localization signal (NLS)

No

Please contact us for more details.

Specificity
Vimentin intermediate filament protein, tested in human, hamster & dog cells

Encoded Nanobody/ VHH
Monoclonal anti-Vimentin single domain antibody (sdAb) fragment

Sequence
With the Vimentin-Chromobody plasmid you receive the sequence information of the Alpaca antibody to Vimentin fused to TagGFP2 or TagRFP, as well as the full vector sequence.

Microscopy techniques
Wide-field epifluorescence microscopy; confocal microscopy

Transfection
Transfection of Chromobody plasmids into mammalian cells can be done with standard DNA-transfection methods, e.g. lipofection (Lipofectamine 2000® from Thermo Fisher Scientific), according to the manufacturer’s protocol for the transfection reagent. Please choose the transfection method that works the best for your cell type.

Storage instructions
Shipped at ambient temperature. Store at -20°C.

Maier J, Traenkle B, Rothbauer U., Visualizing Epithelial-Mesenchymal Transition Using the Chromobody Technology. Cancer Res. 2016 Oct 1;76(19):5592-5596. Epub 2016 Sep 15. Review. PMID: 27634766
Maier J, Traenkle B, Rothbauer U., Real-time analysis of epithelial-mesenchymal transition using fluorescent single-domain antibodies. Sci Rep. 2015 Aug 21;5:13402. doi: 10.1038/srep13402. PMID:  26292717

Are Chromobodies constitutively expressed?

Yes, Chromobody expression is regulated by immediate early promotor CMV. This promotor allows constitutive Chromobody expression.

Do Chromobodies only work in live cells?

Yes, the Chromobody plasmid is only expressed in live cells. Cells should be transfected with the Chromobody plasmid at least overnight to observe the Chromobody location signal. Alternatively, cells can be fixed prior to imaging.
Note: We don't recommend fixation of cells for the Histone-Chromobody.

When should I image my cells after transfection with the Chromobody plasmid?

The Chromobody signal is maintained up to 3 days in the cell. However, this also depends strongly on the cell type.
We recommend to image the cells 16-24 hours after transfection.

Do the Chromobodies diffuse through the cell membrane into growth medium?

No, Chromobodies are small proteins being expressed in the cytosol. They are not secreted into the medium and remain in the cell as long as the cell maintains its plasma membrane integrity.

Are Chromobodies fluorogenic or do they only emit fluorescence when bound to a target?

Chromobodies are chimeric proteins consisting of a VHH fused to a fluorescent protein. They maintain their fluorescence regardless of whether they are bound to a target or not.

Can I amplify the Chromobody plasmid in bacteria?

Yes, the Chromobody plasmids can be propagated in E.coli by standard techniques.

Trace Vimentin dynamics in live cells non-invasively

The transient binding of the Vimentin-Chromobody does not influence cell viability or cell motility and thus offers you the unique possibility to non-invasively monitor the vimentin cytoskeleton in live cells.

Download Whitepaper
Order Free Test-Sample
Email newsletter

Only for research applications, not for diagnostic or therapeutic use!