產(chǎn)品詳情

  • 產(chǎn)品名稱：VimentinChromobody?-TagGFPplasmid

  • 產(chǎn)品型號：
  • 產(chǎn)品廠商：ChromoTek
  • 產(chǎn)品文檔：
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簡單介紹：
VimentinChromobody?-TagGFPplasmid
詳情介紹：
Purpose With the Vimentin-Chromobody? plasmid you can visualize vimentin cytoskeleton in real time in your cell line of interest and analyze epithelial-mesenchymal transition in tumor cell progression.
Brand Chromobody?
Specificity All Chromobody? binding domains are carefully selected to not interfere with endogenous protein function and offer the unique possibility to trace native proteins in living cells.
Characteristics Chromobodies? are a novel species of extremely small intracellular functional antibodies.

Chromobodies? are based on the antigen binding domain (VHH) derived from heavy chain antibodies of camelids genetically fused to a fluorescent protein. As opposed to conventional antibodies, these innovative fluorescent nanoprobes are suitable for real time analyses and can visualize endogenous cellular structures and processes in live cells.
Thus, Chromobodies? are the perfect reagents for cellular research including High-Content Analysis (HCA).

The Chromobody? plasmid allows you to create your own cell lines or transiently express the Chromobody?.
Alternative Name Vimentin
Background 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.
Research Area Cytoskeleton, Lineage Markers, Cell/Tissue Markers, Neural Stem Cell marker
Application Notes Chromobodies? are a novel species of intracellular functional antibodies. Chromobodies? are made by genetically fusing the antigen binding domain (VHH) derived from heavy chain antibodies of camelids to fluorescent proteins, e.g. TagGFP2 or TagRFP.

With ChromoTek's unique Chromobody?-Technology the discrepancy and limitations of antibodies and fluorescent protein tagged reporters can now be overcome. You are now able to study the endogenous proteins of your target of interest in living cells - instead of artificial fluorescent fusion proteins which may behave differently from the real protein or may even be toxic to the cells. In combination with High-Throughput Microscopy and automated image analysis Chromobodies? set up a new class of intracellular biomarkers for High-Content Analysis (HCA). Our proof-of-principle studies include:
  • cell cycle monitoring
  • real time apoptosis studies
  • detection of posttranslational modification


These reagents thus significantly expand both the quality and the quantity of information that can be gathered in HCA.
Comment

The new tool for detection of vimentin cytoskeleton in live cells, the Vimentin-Chromobody?, facilitates real-time analyses of endogenous vimentin in a physiological context. The Vimentin-Chromobody? enables you to investigate dynamic organization of vimentin intermediate filaments and study epithelial-mesenchymal transition.

Features:
Easily transfect your cells with the Vimentin-Chromobody? plasmid
Use Vimentin-Chromobody? as marker of vimentin cytoskeleton
Use Vimentin-Chromobody? to monitor dynamics of vimentin intermediate filaments in live cells
Use Vimentin-Chromobody? to trace epithelial-mesenchymal transition
Vimentin-Chromobody? does not affect cell viability
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

Competition and 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.
Protocol New way to trace the dynamics of vimentin cytoskeleton in live cells
With the Vimentin-Chromobody? plasmid you obtain the sequence of the Alpaca antibody to vimentin fused to TagGFP (Evrogen). Upon transformation, your cells express the Vimentin-Chromobody? and the vimentin filaments are visualized by binding of the fluorescent Chromobody?. The binding does not influence cell viability or cell motility and thus offers you the unique possibility to non-invasively monitor vimentin cytoskeleton in your cells.
Restrictions For Research Use only
Storage -80 °C
Storage Comment Store at -80°C. Short term storage at -20°C.