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H. Environmentally-Sensitive Fluorophore

Solvent-sensitive Fluorescent Probe


1. Toutchkine, A., Kraynov, N. and Hahn, K.: "Solvent-sensitive dyes to report protein conformational changes in living ells", J. Am. Chem. Soc., 125, 4132 (2003)
I-SO-IAA is an environmentally-sensitive fluorophore to proteins, which has merocyanine skeleton and iodoacetamide. The fluorescence quantum yield is sharply dependent on solvent polarity or viscosity, enabling them to report changes of fluorescence intensity in their protein environment. Additionally, it can be excited with visible light, unlike other probes.
[Labeling Procedure]]
1. Preparing Protein Solution
Dissolve Protein at 50~100μmol/L in a suitable buffer such as 10~100mM phosphate, Trisand HEPES at pH 7.0~7.5 and room temperature. Reduce disulfide bonds of the protein with a 10-fold molar excess of DTT.
2. Preparing I-SO-IAA Solution
Dilute I-SO-IAA with DMSO to a final concentration of 10mmol/L (=Stock Solution). Protect it from light as much as possible.
3. Labeling the Protein with I-SO-IAA
Add approximately 2~10 moles of I-SO-IAA for each mole of protein. React for 1 hour at room temperature or overnight at 4°C.

Description Wako Cat. No. (Pkg. Size) Grade Note
I-SO-IAA 093-05371 (5 mg) for Cellbiology Ship and keep at -20˚C in a dark place
1H-Indole-1-propanesulfonic acid, 2-[(2E,4Z)-4-(1,1-dioxide-3-oxobenzo[b]thien-2(3H)-ylidene)-2-butenylidene]-2,3-dihydro-5-[(iodoacetyl) amino]-3,3-dimethyl-,monosodium salt, (2Z)-(9C1)
C27H26IN2NaO7S2 = 704.53
[Fluorescence] λex: 600 nm; λem: around 630 nm
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Wako Product Update No.19 [ page. 16 ]