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Live-cell actin dynamics monitoring kit with fluorescent proteins

This kit is designed to monitor actin dynamics in live cells. The kit includes lentiviral vectors encoding actin fusions to DD-AcGFP1 (green, destabilized) and mCherry (red), and the DD's stabilizing ligand Shield1. DD-AcGFP1-Actin is continuously targeted for degradation in the cell by the proteasomes, unless the cells are cultured in medium containing the stabilizing ligand Shield1. By contrast, mCherry-Actin (which does not contain the DD) has normal stability upon expression, and is constitutively present in the cell.

Adding and removing Shield1 creates a “pulse-chase” like set of conditions which allow you to monitor polymerization of actin monomers as newly synthesized DD-AcGFP1-Actin that is stabilized by Shield1 (green) is integrated into the existing (red) mCherry-Actin actin filament network.

Cat. # Product Size License Quantity Details
631076 Lenti-X™ Actin Dynamics Monitoring Kit Each USD $850.00

License Statement

ID Number  
57 This product is covered by U.S. Patent Nos. 8,173,792 and 9,487,787.
55 cPPT Element. This product and its use are the subject to one or more of the following U.S. Pat. No. 8,093,042. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot disclose information, sell or otherwise transfer this product, its components or materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for any commercial purposes. If the buyer is not willing to accept the limitations of this limited use statement, Takara Bio USA, Inc. is willing to accept return of the product with a full refund. For information on purchasing a license to the DNA-Flap technology for purposes other than research, contact the Transfer of Technology Office, Institut Pasteur, 28 rue du Docteur Roux, 75 724 Paris Cedex 15 (www.pasteur.fr).
69 The DsRed-Monomer, DsRed Express, E2-Crimson and the Fruit Fluorescent Proteins are covered by one or more of the following U.S. Patents: 7,250,298; 7,671,185; 7,910,714; 8,664,471 and 8,679,749.
72 Living Colors Fluorescent Protein Products:

Not-For-Profit Entities: Orders may be placed in the normal manner by contacting your local representative or Takara Bio USA, Inc. Customer Service. Any and all uses of this product will be subject to the terms and conditions of the Non-Commercial Use License Agreement (the “Non-Commercial License”), a copy of which can be found below. As a condition of sale of this product to you, and prior to using this product, you must agree to the terms and conditions of the Non-Commercial License. Under the Non-Commercial License, Takara Bio USA, Inc. grants Not-For-Profit Entities a non-exclusive, non-transferable, non-sublicensable and limited license to use this product for internal, non-commercial scientific research use only. Such license specifically excludes the right to sell or otherwise transfer this product, its components or derivatives thereof to third parties. No modifications to the product may be made without express written permission from Takara Bio USA, Inc. Any other use of this product requires a different license from Takara Bio USA, Inc. For license information, please contact a licensing representative by phone at 650.919.7320 or by e-mail at licensing@takarabio.com.

For-Profit Entities wishing to use this product are required to obtain a license from Takara Bio USA, Inc. For license information, please contact a licensing representative by e-mail at licensing@takarabio.com.

Not-For-Profit Non-Commercial Use License:
A copy of the Lenti-X™ Actin Dynamics Monitoring Kit product License Agreement can be found by clicking here.
44 The DsRed-Monomer and the Fruit Fluorescent Proteins are covered by one or more of the following U.S. Patents: 7,671,185, 7,9110,714 and 8,664,471.
39 AcGFP is covered by U.S. Patent No. 7,432,053.

Use this kit to monitor the highly dynamic behavior of the actin filament system in live cells. The Lenti-X Actin Dynamics Monitoring Kit includes lentiviral vectors encoding actin fusions to DD-AcGFP1 (green, destabilized) and mCherry (red), as well as the DD's stabilizing ligand Shield1. Once your cells have been infected with the included vectors, DD-AcGFP1-Actin is continuously targeted for degradation by the proteasomes, unless the cells are cultured in medium containing the stabilizing ligand Shield1. By contrast, mCherry-Actin (which does not contain the DD) has normal stability and is constitutively present in the cell.

Adding and removing Shield1 creates a "pulse-chase" like set of conditions which allow you to monitor how newly synthesized, Shield-stabilized DD-AcGFP1-Actin (green) is integrated into the existing (red) mCherry-Actin actin filament network.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data

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AcGFP1 is ideal for multicolor and fluorescence microscopy applications

AcGFP1 is ideal for multicolor and fluorescence microscopy applications

AcGFP1 is ideal for multicolor and fluorescence microscopy applications. AcGFP1 and DsRed2 protein fusions were transiently transfected and visualized by fluorescence microscopy. Panel A. pAcGFP1-Mito (mitochondria) and pDsRed2-Nuc (nucleus) in HEK 293 cells. Panel B. pAcGFP1-Golgi (Golgi apparatus) and pDsRed2-Nuc (nucleus) in HEK 293 cells.

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Organelles targeted by the subcellular localization vectors

Organelles targeted by the subcellular localization vectors

Organelles targeted by the subcellular localization vectors.

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Subcellular localization vectors encode fusions of fluorescent proteins with localization signals or subcellular structural proteins, which target the fluorescent protein to a specific organelle or subcellular structure

Subcellular localization vectors encode fusions of fluorescent proteins with localization signals or subcellular structural proteins, which target the fluorescent protein to a specific organelle or subcellular structure

Subcellular localization vectors encode fusions of fluorescent proteins with localization signals or subcellular structural proteins, which target the fluorescent protein to a specific organelle or subcellular structure. The vectors are available in a variety of organelle- and cytoskeleton-targeted color variants.

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Observe actin filament remodeling using the Lenti-X Actin Dynamics Monitoring Kit

Observe actin filament remodeling using the Lenti-X Actin Dynamics Monitoring Kit

Observe actin filament remodeling using the Lenti-X Actin Dynamics Monitoring Kit. Actin filaments are completely remodeled in HeLa cells in less than 1 hr. Panel A. Self-assembly of actin filaments occurs at the plus end of an existing actin filament as monomeric actin is incorporated. Conversely, disassembly occurs at the minus end where actin monomers depolymerize from the filament, causing a continuous rearrangement of the actin filament network. HeLa cells were infected with constructs encoding mCherry human alpha-Actin and DD-AcGPF1 human alpha-Actin. Cells were fixed using 4% paraformaldehyde and imaged with a 40X objective. Fluorescence micrographs were taken 1 hr after addition of Shield1 (Panels E–G) or without Shield1 (Panels B–D). In the absence of Shield1, DD-AcGFP1-Actin was degraded quickly (Panels C & D) despite a normal, mCherry-labeled actin filament network (Panels B & D). In the presence of Shield1, DD-AcGFP1-Actin was stabilized and present in the actin filament network along with mCherry-labeled Actin (Panels E–G). Our results are in agreement with a previous report that the actin filament network rearranges completely in 1 hr in PtK2 epithelial cells, but we used the extremely simple ProteoTuner-based method, rather than the extremely laborious microinjection method.

Overview

  • Live-cell monitoring of actin dynamics
  • Easy-to-use, dual-color assay

More Information

Applications

  • Labeling actin filaments in cells
  • Monitoring actin dynamics in live cells

Additional product information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.

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Takara Bio USA, Inc. provides kits, reagents, instruments, and services that help researchers explore questions about gene discovery, regulation, and function. As a member of the Takara Bio Group, Takara Bio USA is part of a company that holds a leadership position in the global market and is committed to improving the human condition through biotechnology. Our mission is to develop high-quality innovative tools and services to accelerate discovery.

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