The 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.

 

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

 

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.

 

Recommended antibodies

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

WB: Western blotting, IP: Immunoprecipitation

 

References

※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.

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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.
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