多肽标记FITC方法

多肽标记FITC方法

Reagents:
FITC 
Fmoc-e-Ahx-OH

• Couple Fmoc-e-Ahx-OH to the amino terminal of the peptide-resin using standard coupling conditions.

• "De-Fmoc" with piperidine using the standard 20% piperidine procedure.

• Wash resin with DMF (3-4x).

• Swell resin with DCM and drain.

• Prepare solution of 1.1 equivalent of FITC in pyridine/DMF/DCM (12:7:5). Use just enough solution to form a slurry with the resin. Do not use too much solution since the rate of the reaction is proportionate to the concentration of the solution.

• Add the solution prepared in step 2 to the resin.

• Let mix overnight.

• Check the completion of the reaction using ninhydrin test.

• If the coupling of FITC to the amino group is not complete, ninhydrin test will give a blue color. Repeat the coupling with FITC (steps 5-7) if necessary.

• Wash resin with DMF (2x), isopropanol (2x), and DCM (2x).