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].
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.