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EosFP and IrisFP Photoconvertible Fluorescent Proteins

VS-FLP10050 - pmIrisFP, FLAG®-tagged, 10µg

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pmIrisFP is a cloning and expression vector, encoding for the photoactivatable fluorescent protein mIrisFP. mIrisFP combines irreversible photoconversion from a green- to a red-emitting form with reversible photo-switching between a fluorescent and a non-fluorescent state in both forms. The protein is an advanced monomeric variant of EosFP that differs from the monomeric mEosFP by four additional mutations: A69V, K145I, F173S and Y189A. 
 
mIrisFP has excellent properties as a genetically encoded fluorescence marker. It matures with a half-life of 14 min at 37°C, faster than many engineered fluorescent proteins in vitro. In HEK293 cells, mIrisFP green fluorescence appears 15h after transfection, similar to EGFP (13h). mIrisFP fluorescence can be switched on and off reversibly, but also allows for the non-reversible photoconversion from green to red. The dual photoactivation capability of mIrisFP offers enhanced flexibilitiy and enables the combination of pulse-chase experiments with super-resolution imaging. 
 
See below for the optical parameters of mIrisFP:
 

Parameter

Before photoconversion

After photoconversion

Excitation / Emission

485 nm / 516 nm

546 nm / 578 nm

Extinction coefficient

47'000M-1cm-1

33'000M-1cm-1

Fluorescence Quantum Yield

0.54

0.59

Brightness x 1000

25.5

19.3

Switching half-time off / on

8.8 s / 6.4 s*

16.0 s / 4.9 s**

Relaxation half-time

53 min

24 min

Detected photons per burst

176

354

*off-switching: 473 nm light, 30 mW cm-2, on-switching: 405 nm light: 4.5 mW cm-2
**off-switching: 561 nm light, 6 mW cm-2, on-switching: 473 nm light: 0.5 mW cm-2
 

Features

  • Expression vector for the fluorescent protein mIrisFP
  • Ready-to-use construct for cell tracking
  • Green-to-red photoconvertible and photoactivatable
  • Permanent, bright and fast
  • Monomeric; low tendency for dimer formation
  • Suitable for pulse-chase experiments, measurements of kon- and koff- rates, and high-resolution fluorescence microscopy such as PALM
  • Source vector for creating fusions with signal peptides
 

Applications

  • Selective marking of cells, subcellular compartments and fusion proteins by photoconversion
  • Localization studies of fusion proteins with mIrisFP
  • Pulse-chase experiments and superresolution imaging with mIrisFP
 

Contents

  • pmIrisFP, FLAG-tagged, lyophilized DNA, 10 µg
 

Literature

 

Publications

  • Fuchs, J., Boehme, S., Oswald, F., Hedde, P.N., Krause, M, Wiedenmann, J., and Nienhaus, G.U. (2010). Imaging Protein Movements in Live Cells with Super-resolution Using mIrisFP. Nature Methods 7, 627 – 630.
 
  • McKinney, S.A., Murphy, C.S., Hazelwood, K.L., Davidson, M.W. & Looger, L.L. (2009). A bright and photostable photoconvertible fluorescent protein. Nature Methods 6, 131-133.