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德国ChromoTek® Vimentin Chromobody®-TagGFP plasmid!北京乐博生物,授权代理
lebo / 2016-05-11
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ChromoTek® Vimentin Chromobody®-TagGFP plasmid; Product Code ‘vcg’

Product Information Page

1. Background

Vimentin: The type III intermediate filament protein Vimentin is the major cytoskeletal component of mesenchymal cells. It is expressed in a highly developmentally-regulated fashion. Therefore vimentin has become an important biomarker of mesenchymally-derived cells or cells undergoing an epithelial-to-mesenchymal transition (EMT) during both normal development and metastatic progression, highly dynamic cellular processes.

Chromobodies: ChromoTek has developed Chromobodies®, which visualize endogenous proteins in live cells and at real time, to overcome limitations of the current protein visualization methods antibody staining and fluorescent fusion protein expression:

· Antibodies are valuable tools for the visualization of endogenous cellular components, however this procedure requires cell fixation restricting their applicability to end-point measurements. The traditional antibodies’ use in living cells even as fragments is limited because of the inefficient folding and assembly of their variable heavy and light chains.

· Fluorescent fusion proteins can easily be overexpressed in cells and provide information on protein localization and dynamics in living cells, but endogenous proteins remain invisible. Fusion protein (over-) expression may also be cytotoxic and may introduce artificial effects.

2. Product Description

The Vimentin Chromobody is a functional intracellular antibody (so called “nanobody”/VHH derived from Alpaca), which is genetically fused to Green Fluorescent Protein (TagGFP2 only!) for real time analysis of endogenous vimentin in live cells. The Vimentin Chromobody is available as plasmid for mammalian cell transfection. It is sold as plasmid in quantities of 20 µg using product code “vcg” at standard Chromobody pricing. The Vimentin Chromobody-TagGFP plasmid is shipped at room temperature and is accompanied by a vector map. Lentiviral particles and stable cell lines are available on request as custom services. Please inquire. Note: A genetic fusion to tagRFP is not available.

3. Positioning Statement

The Vimentin Chromobody is a proprietary, ground breaking tool for detection of endogenous vimentin in real-time in live cells. It is designed to overcome limitations of protein analysis in cells using antibody staining and fusion protein expression.

4. Value Proposition

The Vimentin Chromobody is a unique tool for visualization of endogenous vimentin in real-time in live cells. It has been carefully tested to not interfere with the biological functions of vimentin.

· Dynamic live-cell analysis, (but also end-point assays if needed)*

· Fast no-wash assay (compared to antibody staining)*

· Visualization of endogenous vimentin

· No vimentin overexpression, which would in fact induce EMT phenotype

· No cytotoxicity and artificial effects

*Main advantages of the Vimentin Chromobody.

5. Targeted Markets & Customers

The targeted markets comprise oncology research markets in both academia and pharma. Here, cell biologists working in cancer and/or having the need to visualize vimentin as a biomarker in live cells. In pharma, small molecule screening departments in drug discovery divisions (secondary screens) are addressed.

6. 
Customers’ Applications

· Cell biological, i.e. microscopic/HCA analysis of cells, tissues, whole organs (sections) and organisms (sections)

· Examination of cancer progression visualizing epithelial-mesenchymal transition (Vimentin)

· Cancer research including but not limited to: mesenchyme, colon cancer, certain upper gastrointestinal pathologies, and hormone positive breast cancers.

· Investigation of small molecules in (secondary) screens – Vimentin is a biomarker

7. Competition & Competing Technologies

· Traditional antibody staining is the major technology to visualize vimentin. Hence there are hundreds of anti-vimentin antibodies commercially available. To select a proper, validated antibody is challenging and time consuming. In addition the staining is end-point only and the protocol for fixation and staining with primary and secondary is time consuming (3h vs. 30 min when using the Vimentin Chromobody).

· Fusion Protein Expression - genome engineered visualization tag/fusion protein under endogenous promotor control (using e.g. TALEN or CRISPR/Cas) only. It is understood that these procedures are still challenging not as straight forward as being reported elsewhere. Note: Heterologous expression would result in overexpression of vimentin and thus induce the EMT phenotype, making its investigation impossible.

8. Sales Strategy

We recommend to promote the ChromoTek web-based reference database for latest publications on Chromobodies. Also the ChromoTek senior scientists are available for technical advice and to assist in the sales process. NB: a EULA (End User License Agreement) is required to be accepted by the customer as a prerequisite of the sale prior to shipment. ChromoTek will contact you accordingly on any order you may place for Chromobodies.

9. Key Words

Vimentin, epithelial (to) mesenchymal transition, EMT, biomarker, intracellular antibody/nanobody, Chromobody, intrabody, live cell imaging, real-time visualization, cancer, oncology, small molecule screens

 

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