多肽标记FITC方法

上海沪震生物科技有限公司

2017/12/23 16:49
TA的动态

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).


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