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Midoriishi-Cyan

  • FRET donor with highly fluorescent quantum yield
  • Cell, subcellular structure, or protein labeling

 Expressiom of Midoriishi-Cyan in HeLa cellsThe fluorescent protein Midoriishi-Cyan gene was isolated from the stony coral Acropora sp.(Midori-ishi in Japanese).

CoralHue® Midoriishi-Cyan 1 (MiCy1) absorbs light maximally at 472 nm and emits cyan light at 495 nm. It rapidly matures to form a bright dimeric complex. So it can be used to mark individual cells or to report gene expression without the problem of protein aggregation. CoralHue® mMiCy1 is a monomeric version of MiCy1, and can be used to label proteins or subcellular structures. These fluorescent proteins are suitable as donor for fluorescence resonance energy transfer (FRET) because of highly fluorescent quantum yield.


 

Fluorescent properties

MiCy1


mMiCy1




Midoriishi-Cyan1 spectrum data files (text files)
Midoriishi-Cyan1 excitation (8K)  Midoriishi-Cyan1 emission (16K)
mMidoriishi-Cyan1 excitation (8K)  mMidoriishi-Cyan1 emission (4K)

Note: The file is in a tab-delimited text format. It contains values of the wavelength (0.5nm spacing) and brightness (fluorescence intensity peak value normalized to 1). Use a spreadsheet program to create a spectrum that will help you in choosing the appropriate excitation filter, dichroic mirror and fluorescence filter.

CHARACTERISTIC CoralHue® MiCy1 CoralHue®mMiCy1
Oligomerization dimer monomer
Number of amino acid 232 232
Excit./Emiss. maxima (nm) 472/495 470/496
Molar extinction coefficient (M-1cm-1) 27,250 (472 nm) 22,150 (470 nm)
Fluorescence quantum yield 0.90 0.70
Brightness*1 24.5 15.5
pH sensitivity pKa=6.6 pKa=7.0
Cytotoxicity*2 No No

*1Brightness: Molar Extinction Coefficient ×Fluorescence Quantum Yield / 1000
*2Toxicity when expressed in HeLa cells

 

Performance and use

Monitoring of caspase-3 activity during apoptosis

MiCy and Kusabira-Orange (KO) is a good donor/acceptor pair for FRET (fluorescence resonance energy transfer). FRET based caspase3 monitoring probe is constructed using MiCy and KO, connected by a peptide containing the caspase3 cleavage sequence, DEVD (Asp-Glu-Val-Asp). The activation of caspase3 leads to eliminate FRET by cleavage of DEVD sequence.

Cell: HeLa
Filter, mirror: DM 465HQ, Ex BP440-460HQ, Em BA475-540HQ(cyan), BA555-600HQ(orange)
Lamp: Xe lamp (75W)
Lens: UApo/340 N.A. 1.35
Apoptosis induction: 500 ng/ml of anti-Fas antibody (CH-11; Medical & Biological Laboratories Co.,Ltd.) and 10 μg/ml cycloheximide at 36h post-transfection.

AG resistant to acidity


 

Recommended antibodies

CoralHue® Midori-ishi Cyan can be recognized using antibodies as shown below.

  • Anti-MiCy for WB, Code No. M130-3M
  • Anti-MiCy for IP, Code No. M116-3M

WB: Western blotting, IP: Immunoprecipitation

 

References

  • Karasawa S, Araki T, Nagai T, Mizuno H, Miyawaki A, Cyan-emitting and orange-emitting fluorescent proteins as a donor/acceptor pair for fluorescence resonance energy transfer; Biochem J. 381, 307-12(2004) PMID:15065984

CoralHue® fluorescent proteins were co-developed with the Laboratory for Cell Function and Dynamics, the Advanced Technology Development Center, the Brain Science Institute, RIKEN. MBL possesses the license and deals in the products.


Please send us a completed and signed License Acknowledgement.

Please note that the product will be shipped after we receive a completed and signed License Acknowledgement (PDF file on the right). Prior to shipping, our staff may contact you to verify the intended use of the product. Please also note that a separate agreement or a contract may be required depending on the intended use.
The product you have ordered is sold for research purpose only. It is not intended for industrial or clinical use, and shall not be used for any purposes other than research. You are also asked not to hand over or resell the product to a third party.
If you belong to a profit organization or a business corporation, or wish to use the product for profit or commercial purposes, please contact us.


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