Universal C-terminal Protein Labeling with Coumarin
For in vitro use only!
Shipping: shipped on blue ice
Storage Conditions: store at -20 °C
avoid freeze/thaw cycles, store dark
Shelf Life: 12 months
Spectroscopic Properties: λexc 332 nm; λem 410 nm (Keto-Coumarin)
Coumarin Protein-C-labeling Kit provides a highly efficient and easy-to-handle tool for the C-terminal modification of proteins based on a chemoselective oxime ligation. The rapid reaction of oxyamino modified proteins with ketones under mild condition is potentially beneficial for many biological applications. The kit contains Keto-Coumarin as fluorescent label.
pTWIN Vector (red cap)
5 μg modified pTWIN vector
Bisoxyamine (blue cap)
111 μl 0.9 M in Reaction Buffer
MESNA (yellow cap)
35 mg (Sodium 2-mercaptoethanesulfonate)
Keto-Coumarin (purple cap)
12.5 μl 100 mM
To be provided:
25 mM NaH2PO4 (pH 7.5), 0.5 M NaCl
50 mM NaH2PO4 (pH 8.0), 0.3 M NaCl
50 mM NaH2PO4 (pH 8.0), 0.3 M NaCl, 0.5 M imidazole
30 mM NaH2PO4 (pH 7.5), 50 mM NaCl, 10 mM MESNA
30 mM NaH2PO4 (pH 7.5), 50 mM NaCl
30 mM NaH2PO4 (pH 7.5), 50 mM NaCl, 2 mM DTE
30 mM NaH2PO4 (pH 7.0), 50 mM NaCl, 2 mM DTE
50 mM NaAc (pH 5.5), 50 mM NaCl, 2 mM DTE
Site-specific protein modification can facilitate the characterization of protein functions both in biochemical and in cellular investigations. Although many chemical reactions are applicable in principle, methods for the site-specific modification of proteins remain in high demand, and there is a requirement for readily available ligation reagents and mild reaction conditions. Oxime-based reactions have found wide application in the conjugation of biomolecules on account of the absence of oxyamino groups in proteins and their orthogonal reactivity with ketones to give stable oximes.
The oxyamine-ketone bioorthogonal reaction has been exploited in protein modification mainly by means of incorporating ketone groups into proteins by various chemical, enzymatic, and molecular biological methods. To expand the application of this efficient methodology to protein ligation, a simple and general method to incorporate oxyamino groups into proteins has been developed.
Protein Preparation with C-terminal-thioesther
Protein C-terminal Labeling