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ChromoTek - Specific Application Areas
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Specific Application and Product Information on the ChromoTek Range of Products
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Information on Specific Application Areas for ChromoTek products can be found below. For full information on any ChromoTek products, please visit the ChromoTek website.
Specific Product/Application Areas:
Go to Technical Overview - NanoTrap Technology Technical Overview - NanoTrap Technology
Go to GFP-Trap Products GFP-Trap Products
Go to RFP-Trap Products RFP-Trap Products
Go to GFP-multiTrap GFP-multiTrap
Go to GFP-Booster GFP-Booster
Go to Chromobody Products Chromobodies
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Technical Overview - NanoTrap Technology
The green fluorescent protein (GFP) was originally isolated from the jellyfish Aequora victoria. GFP absorbs blue light and emits green light with an emission peak of ~509 nm. In molecular and cell biology green fluorescent proteins and spectral derivates thereof are the most popular tools to study gene expression, protein localisation and dynamics in vivo.

Technology:
Antibodies are extremely powerful tools in biomedical research and are large complex molecules (~150 kDa) consisting of two heavy and two light chains. Due to their complex structure, the use of antibodies can be limited and hindered by batch-to-batch variations.

Camelidae (camels, dromedaries, llamas and alpacas) possess functional antibodies devoid of light chains, so-called heavy chain antibodies (hcAbs). hcAbs recognise and bind their antigens via a single variable domain fragment (VHH). These VHH domains are the smallest intact antigen binding fragments (~13 kDa).
 
 
The Nano-Trap offers a high quality, single domain GFP binding protein coupled to a matrix for biochemical studies. ChromoTek's GFP-Trap® and RFP-Trap® and now well established a novel, high quality system for the fast, reliable and efficient one-step isolation of fluorescent fusion proteins and their interacting factors.
Comparison of GFP-Trap® with conventional mono- and poly-clonal antibodies:
Immunoprecipitations (IP) of GFP from protein extracts of GFP-producing human cells. Input (I), non-bound (FT) and bound (B) fractions were separated by SDS-PAGE followed by Coomassie staining and Western Blotting. (hc) heavy chain, (lc) light chain of conventional antibodies.
Comparison of GFP-Trap®_A and GFP-Trap®_M:
Left (IP): Pulldown of GFP with GFP-Trap®_A and GFP-Trap®_M from 293T cell extracts. Input (I) and bound (B) fractions were separated by SDS-PAGE followed by Coomassie staining.

Right (Co-IP): Pulldown of GFP-PCNA with GFP-Trap®_A and GFP-Trap®_M from 293T cell extracts. Other bands: potential interaction partners of PCNA.
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Products Product Description
GFP-Trap Products
 
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GFP-Trap - a versatile tool for biochemical and cell biological analysis of GFP-tagged fusion proteins
 
The GFP-Trap® specifically binds to most common GFP derivatives:
  eCFP, CFP, mCerulean
  eGFP, wtGFP, GFP S65T, AcGFP, TagGFP, tagGFP2, sfGFP, pHluorin, Clover, GFP Envy
  eYFP, YFP, Venus, Citrine
 
GFP-Trap® is a novel high quality system for the fast, reliable and efficient one-step isolation of fluorescent fusion proteins and their interacting factors. GFP-Trap® can be used for a number of important biochemical analyses:
  Pulldown
  Immunoprecipitation (IP)/Co-IP
  Mass spectrometry
  Enzyme activity measurements
  ChIP analysis
 
GFP-Trap® is extremely stable (up to 70°C, remains functional in 2M NaCl or 0.5% SDS) and has a high binding affinity (dissociation constant in the sub-nanomolar range). GFP-Trap® specifically binds to eGFP derivatives such as GFPS65T, YFP or eYFP while and shows no detectable binding to proteins derived from DsRed.
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RFP-Trap Products
 
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RFP-Trap - a versatile tool for biochemical and cell biological analysis of RFP-tagged fusion proteins
 
The RFP-Trap® specifically binds to most common RFP derivatives:
  mRFP, mCherry, mRFPruby, tdTomato, mPlum, tagRFP, mKate2, DsRed
 
RFP-Trap® is a novel high quality system for the fast, reliable and efficient one-step isolation of fluorescent fusion proteins and their interacting factors. RFP-Trap® can be used for a number of important biochemical analyses:
  Pulldown
  Immunoprecipitation (IP)/Co-IP
  Mass spectrometry
  Enzyme activity measurements
 
RFP-Trap® efficiently pulls down various monomeric red fluorescent proteins derived from DsRed, e.g. mRFP1, mCherry, mOrange, mPlum but also mRuby and RFP-tagged fusion proteins. No cross-reaction to GFP derivatives is observed.
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GFP-multiTrap
 
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GFP-multiTrap - take advantage of the proven efficiency of GFP-Trap in a convenient 96-multiwell format
 
GFP-Trap® is immobilised directly onto the wells, thus eliminating time consuming centrugation of individual tubes, making it easy to quickly test for GFP fusion proteins for peptide, protein, DNA or RNA binding.

The 96-well format allows for the rapid quantification of the the washed and bound fractions of GFP fusion proteins and fluorescently labelled binding substrates when used with fluorescence scanners and plate readers.
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GFP-Booster
 
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GFP-Booster "Nanoscopy" Increases Signal Intensity
 
GFP signal intensities at physiological expression levels are usually too low for super-resolution microscopy. ChromoTek's GFP-Booster enables the use of existing expression constructs and cell lines for super-resolution microscopy. With the GFP-Booster it is now possible for the first time to study the protein machinery involved in the process, the last step of cell division. The fluorescence enhancement achieved by the GFP-Booster enables super-resolution microscopy to resolve the 3D distribution of GFP-tagged proteins and provides novel insights into the cell division process.
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Chromobodies
 
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Chromobodies express novel intracellular antibodies
 
The new Chromobodies® Plasmids express 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 (e.g.TagGFP or TagRFP from Evogen). As opposed to conventional antibodies, these innovative fluorescent nanoprobes are suitable for real time analyses and can visualise endogenous cellular structures and processes in living cells. All Chromobody binding domains are carefully selected to not interfere with endogenous protein functions. The transient binding does not influence cell viability or motility and thus offers a unique opportunity to observe native proteins within the cell. Chromobody plasmids allow you to create your own cell lines or transiently express the Chromobody. Thus, Chromobodies are the perfect reagents for cellular research including High-Content Analysis (HCA).   
   
Five Chromobody Plasmids types are available:  
  Actin-Chromobody plasmid: A new tool for detection of actin in live cells, the Actin-Chromobody, enables the monitoring of the assembly/ disassembly of cytoskeleton in real time..
  Cell Cycle Chromobody plasmid: A powerful Cell Cycle Chromobody  which enables screening of compounds such as cancer drugs for effects on the cell cycle and toxicity in one experiment. This HCA assay can be used in early drug development and validation as well as in basic research.
  Dnmt1-Chromobody plasmid: Trace dynamic changes during the cell cycle in real time witht the Dnmt1-Chomobody. Monitor the distribution of an endogenous cell cycle marker protein - unlike with fluorescent fusion proteins there are no overexpression artifacts or cytotoxic effects
  Lamin-Chromobody plasmid: Visualize the nuclear lamina without interfering with its cellular functions. Monitor the nuclear integrity and morphology during live cell microscopy as an indicator for cellular health and viability. Use the Lamin-Chromobody as a biosensor for real-time assays of mitosis or apoptosis.
  PARP1-Chromobody plasmid: A new tool for detection of PARP1 in live cells, the PARP1-Chromobody, enables the monitoring of the recruitment of PARP1 to DNA damage after microirradiation in real time.
 
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