rLV-Ubi-LifeAct Lentiviral Vectors

£758.00

Key Features

An actin marker used to visualize F-actin in living cells after lentiviral transduction. Especially suited for the generation of stable cell lines.

  • Brilliant visualization of F-actin in live cell imaging – perfect imaging of cytoskeletal organization and cellular dynamics
  • No interference with cytoskeletal dynamics – unrestricted actin functionality
  • Excellent signal-to-noise ratio
  • Easy generation of stable cell lines
  • Effective for dividing and non-dividing cells
  • Lentiviral vector, ready to use
  • Ubi (ubiquitin) promotor
  • Concentration / Volume – 1 x 10^7 TU/ml, 100 µl

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Description

Applications:

  • Cytoskeleton organization:
    – Non-toxic actin visualization
    – Fluorescence staining
  • Cytoskeletal dynamics:
    – Actin dynamics in cellular processes
    Live cell imagingand microscopy

Technical Features:

  • Superb biocompatibility – non-toxic staining of living samples
  • Easy generation of stable cell lines
  • For in vitro and in vivo applications

Specifications:

Concentration 1 x 107 TU / ml
Storage -80 °C
TagGFP2 (Exmax / Emmax) 483 / 506 nm
TagRFP (Exmax / Emmax) 555 / 584 nm

 

NOTE:
Please note that you will be working with a product containing infectious virus. Follow the recommended NIH guidelines for all materials containing BSL–2 organisms.

 

LifeAct, a 17-amino acid peptide, is derived from a protein from Saccharomyces cerevisiae. LifeAct stains filamentous actin (F-actin) structures in living or fixed eukaryotic cells and tissues. In contrast to GFP-actin and its alternatives, such as actin-binding proteins, LifeAct does not interfere with actin dynamics in vitro and in vivo, as shown in several cell types and model organisms.*

Lentiviral vectors derived from the human immunodeficiency virus (HIV-1) have become major tools for gene delivery into mammalian cells. The most advantageous feature of lentiviral vectors is the ability to mediate efficient transduction, integration, and long-term expression into dividing and non-dividing cells, both in vitro and in vivo.

The pseudotyped envelope with vesicular stomatitis virus envelope G (VSV-G) protein broadens the target cell range. Lentiviral vectors have now been shown to deliver genes to neurons, lymphocytes and macrophages. Previous retrovirus vectors could not be used for these cell types. Moreover, lentiviral vectors have proven to be effective in transducing brain, liver, muscle, and retina in vivo without any toxicity effects or immune responses.

The provided lentivirus is a VSV-G-pseudotyped pantropic virus, which is capable of infecting both dividing and non-dividing cells, while stably integrating into the cells’ genome.

After the transduction of cells with rLV-LifeAct, F-actin is visualized using the fluorescence markers TagGFP2 or TagRFP**. With this method, transduction efficiencies of up to 100% can be attained, even in difficult-to-transfect cell types like primary cells (e.g., neuronal cells). This virus can be used to generate stable cell lines, and stable clones can be selected by fluorescence sorting.

LifeAct plasmidsmay be used in many cell types when a transient or stable expression of LifeAct is needed. In this case, researchers can choose between two different promoters: CMV (cytomegalovirus) or CAG (modified chicken β-actin).

Adenoviral vectors are available for difficult-to-transfect cells, or when up to 100 % transgene expressing cells are needed. In this case, it is also possible to choose between the two promoters previously mentioned. These vectors are suitable for applications where only the transient expression of LifeAct-TagGFP2 or LifeAct-TagRFP is needed.

Method Comparison: Using LifeAct for F-Actin Visualization in Living Cells

Method Suitable for 1
Cell lines
+++
+++
+++
+++
Primary cells
+
++
+++
+++
Non-mammalian cells
+
Transfection Stability
Transient
Yes
Yes
Yes
No
Stable
Yes
No
No
Yes
Protocol and Analysis
Protocol simplicity
+++
++
+++
+++
LifeAct readout start
1–2 days
few minutes
1–2 days
1–2 days
LIfeAct readout duration
several days
(infinity if stable)
up to 1 day
several days
several days
(infinity if stable)
Signal intensity
+++
+
+++
+++
Extra Benefits
Prevention of overexpression artefacts
+++
+
+
Prevention of viral response
+++
+++
Biocompatibility of transfer method
+
+++
++
++
Actin functionality
+++
+++
+++
+++
Safety Consideration
Biosafety Level
BSL1
BSL2
BSL2

1 Not all of these combinations have been tested in all indicated cell types. We encourage you to use this table as a guideline to understand how the LifeAct system can be optimally applied.


* Original LifeAct publication in Nature Methods:
Riedl J, et al. Lifeact – a versatile marker for the visualization of F-actin. Nature Methods 5, 605-607 (2008)

** Information about TagGFP2 and TagRFP at www.evrogen.com

LifeAct is a registered trademark of ibidi GmbH.

For license and product use information please refer to www.ibidi.com, Legal Notes.

Additional information

Tag

TagGFP2, TagRFP