Fl-DIBO

Fl-DIBO (fluorogenic dibenzocyclooctyne) is a selective and high sensitivity fluorescent probe to azide compounds. Fl-DIBO can react rapidly with azide compounds to form new highly fluorescent products with a maximum emission wavelength of 469 nm and excitation wavelength of 363 nm. Fl-DIBO can be used to label diazo-tagged proteins without detectable background signal interference[1][2].

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Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs). Labeling diazo-labeled proteins[2][3]: 1. Resuspend the protein with 1×PBS (pH 7.4) and incubate with NHS-activated ester (25 mM in DMSO) overnight at room temperature. 2. Labeled proteins are co-incubated with Fl-DIBO (25-500 μM) at 37 °C for 18 h. 3. Protein SDS-PAGE gel analysis and in-gel fluorescence imaging analysis with λex=365 nm, λem=480 nm.

[1]. Ke Wang, et al. A metal-free turn-on fluorescent probe for the fast and sensitive detection of inorganic azides. Bioorg Med Chem Lett. 2016 Apr 1;26(7):1651-4.

[2]. Frédéric Friscourt, et al. Fluorogenic Strain-Promoted Alkyne-Diazo Cycloadditions. Chemistry. 2015 Sep 28;21(40):13996-4001. doi: 10.1002/chem.201502242. Epub 2015 Aug 18.

[3]. Camille Favre, et al. Sydnone Reporters for Highly Fluorogenic Copper-Free Click Ligations. J Org Chem. 2018 Feb 16;83(4):2058-2066.