CuAAC Cell Reaction Buffer Kit (BTTAA based)

The set-up of a CuAAC reaction is based on the following general three-step procedure: Prepare a mix of Alkyne- and Azide functionalized molecules in an appropriate reaction buffer. Prepare a CuSO4: Cu (I) -ligand premix, add it to the Azide- Alkyne solution and mix briefly. Add Na-Ascorbate as reduction reagent at last to start the reaction. The CuAAC Cell Reaction Buffer Kit (BTTAA based) provides sufficient amounts to perform 50 CuAAC experiments of 500 μl each using 2 mM CuSO4, 10 mM BTTAA and 100 mM Na-Ascorbate in 100 mM Na-Phosphate reaction buffer. We provide a general protocol for labeling of fixed and permeabilized cells containing metabolically functionalized Alkyne- or Azide-modified biomolecules. However, individual optimization might be required for different CUAAC labeling experiments as well as for critical reaction parameter e.g. final CuSO4 concentration, CuSO4:BTTAA ratio, detection reagent concentration.