Actin-Chromobody

Live visualization of actin dynamics with Actin-Chromobody

The Actin-Chromobody® for monitoring assembly/ disassembly of the actin cytoskeleton in live cells in real-time.

The Actin-Chromobody is a chimeric protein for detection of microfilaments. It consists of a fluorescent protein fused to a VHH. The Actin-Chromobody visualizes actin filaments without notable interference with its dynamics s (Melak et al. 2017).

Specificity
Actin microfilaments in human, rodent and plant cells, zebrafish, Toxoplasma gondii

Applications
Live cell imaging
Monitoring actin dynamics
Actin as  biomarker or control in secondary screens

Actin-Chromobody products
Actin-Chromobody plasmids

Product Size Code Price Buy
Product Actin-Chromobody® plasmid (TagGFP2) Size 20 µg Code acg Price $ 999
Buy +
Product Actin-Chromobody® plasmid (TagRFP) Size 20 µg Code acr Price $ 999
Buy +
Product Nuclear Actin-Chromobody® plasmid (TagGFP2) Size 20 µg Code acg-n Price $ 999
Buy +
Product Actin Chromobody cell lines Size Code transduced in collaboration with NMI Tuebingen Price Please inquire

Are Chromobodies constitutively expressed?

Yes, Chromobodies expression is regulated by immediate early promotor CMV. This promotor allows constitutive Chromobody expression.

Do Chromobodies only work in live cells?

Yes, the Chromobody plasmid is only expressed in live cells. Cells should be transfected with the Chromobody plasmid at least over night to observe the Chromobody location signal. Alternatively, cells can be fixed prior to imaging.
Note: We don't recommend fixation of cells for the Histone-Chromobody.

When should I image my cells after transfection with the Chromobody plasmid?

The Chromobody signal is maintained up to 3 days in the cell. However, this also depends strongly on the cell type.
We recommend to image the cells 16-24 hours after transfection.

Do the Chromobodies diffuse through the cell membrane into growth medium?

No, Chromobodies are small proteins being expressed in the cytosol. They are not secreted into the medium and remain in the cell as long as the cell maintains its plasma membrane integrity.

Are Chromobodies fluorogenic or do they only emit fluorescence when bound to a target?

Chromobodies are chimeric proteins consisting of a VHH fused to a fluorescent protein. They maintain their fluorescence regardless of whether they are bound to a target or not.

Can I amplify the Chromobody plasmid in bacteria?

Yes, the Chromobody plasmids can be propagated in E.coli by standard techniques.

Benefits

The Actin Chromobody allows imaging of actin dynamics in real-time in living cells.

  • Easily transfect your cells and non-invasively visualize actin cytoskeleton in real-time
  • Target and trace microfilaments in your cell line of interest
  • Use Actin-Chromobody as a marker of cellular morphology and as a probe for cellular motility
  • Actin-Chromobody has no notable effect cell viability or migration  

Comparison of Actin visualization probes

Overview of current tools to visualize actin filaments

 

Phalloidin

LifeAct

Actin-Chromobody

Utropin

F-tractin

SiR-actin

GFP-actin

Origin

 
  • Amanita phalloides
 
 
  • Saccharomyces cerevisiae
 
 
  • Vicugna pacos
 
 
  • Homo sapiens
 
 
  • Rattus norvegicus
 
 
  • Jaspis johnstoni
 
 
  • various
 

Description

 
  • Phallotoxin
 
 
  • Amino acids 1-17 of Abp140
 
 
  • Anti-actin-nanobody
 
 
  • Amino acids 1-261 of Utrophin
 
 
  • Amino acids 10-52 of ITPKA
 
 
  • SiR and desbromodes-methyl-jasplakinolide
 
 
  • β-actin fusion protein with GFP
 

Applicability

 
  • Gold standard marker for F-actin in fixed samples
 
 
  • Live-cell imaging of endogenous actin
 
 
  • Live-cell imaging of endogenous actin
  • Super-resolution microscopy
 
 
  • Live-cell imaging of endogenous actin
 
 
  • Live-cell imaging of endogenous actin
 
 
  • Live-cell imaging of endogenous actin
  • Super-resolution microscopy
 
 
  • FRAP
  • Live-cell imaging
 

Advantages

 
  • Excellent specificity for F-actin
 
 
  • Widely used in the scientific community
  • Small probe size
 
 
  • Minimal effect on actin dynamics
  • Small probe size
 
 
  • No binding of G-actin
 
 
  • No binding of G-actin
  • Small probe size
 
 
  • No transfection required
 
 
  • Direct labeling and incorporation into native filaments
 

Potential Pitfalls

 
  • Might alter actin dynamics
  • Might stabilize F-actin
 
 
  • Might alter actin dynamics
  • Binding of G-actin
 
 
  • Binding of G-actin

 

 
  • Might alter actin dynamics
  • Might stabilize F-actin
 
 
  • Might alter actin dynamics
  • Might stabilize F-actin
 
 
  • Might alter actin dynamics
  • Might stabilize F-actin
 
 
  • Ectopic actin expression
  • High background
 

Adopted and modified from Melak et al. (2017). Product names may be covered by registered trademarks of corresponding corporations.

Actin-Chromobody Plasmid (TagGFP2)

Mammalian expression vector encoding the cytoskeleton marker Actin-VHH fused to the green fluorescent protein TagGFP2 (Evrogen)

Actin-Chromobody Plasmid (TagRFP)

Mammalian expression vector encoding the cytoskeleton marker Actin-VHH fused to the red fluorescent protein TagRFP (Evrogen)

Nuclear Actin-Chromobody Plasmid (TagGFP2)

Mammalian expression vector encoding the cytoskeleton marker Actin-­VHH fused to TagGFP2 (Evrogen) and a nuclear localization signal (NLS)

Lentiviral Actin-Chromobody® cell lines

(generated in collaboration with NMI Tuebingen)

cell line fluorescent protein color
A549 green
HepG2 green
HeLa green
Ins-1 green
MDA-MB-231 green


Please inquire for more information!

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Only for research applications, not for diagnostic or therapeutic use!