Materials and Reagents

1. Dichloromethane (DCM)
   
2. Dimetylformamide (DMF) (Fisher Science, catalog number: D119-4 )
   
3. Trifluoroacetic acid (TFA) (ACRO, catalog number: 13972 )
   
4. Thioanisole (TIS) (Sigma-Aldrich,  233781 )
   
5. Ethanedithiol (EDT) (TCI, catalog number: E0032 )
   
6. 2-(1H-Benzotriazol-1-yl)-1,1,3,3-tertramethyluronumhexafluorophosphate (HBTU) (Anaspec, catalog number: AS-21001 )
   
7. N,N-diisopropylethylamine (DIEA) (Santa Cruz Biotech, catalog number: sc-215490 )
   
8. Piperidine (biotech. grade, ≥99.5%) (Sigma-Aldrich, catalog number:  571261 )
   
9. Dithiothriotol (DTT)
   
10. Pyridine
    
11. Potassium cyanide
    
12. Ninhydrin
    
13. Fmoc protected amino acids:
    Fmoc-Ile-OH, Fmoc-Glu (OtBu)-OH, Fmoc-Met-OH, Fmoc-Leu-OH, Fmoc-Lys (Boc)-OH, Fmoc-Arg (Pbf)-OH, Fmoc-Asn (Trt)-OH, Fmoc-Ser (tBu)-OH, Fmoc-Ala-OH, Fmoc-His (Trt)-OH, Fmoc-Gln (Trt)-OH, Fmoc-Val-OH, Fmoc-Phe-OH, Fmoc-Thr(tBu)-OH, Fmoc-Pro-OH and Fmoc-Asp (OtBu)-OH, Fmoc-Gly-OH, Fmoc-Cys(Trt)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Trp(Boc)-OH.
    
14. Rink Amide Resin (0.66 mmol/g) (AAPPTec, catalog number: RRZ001 or equivalent)
    
15. C-18 column (Vydac, catalog number: 218TP510 or equivalent)
    All reagents are ACS grade or synthesis grade
    
16. Anhydrous diethyl ether
    

Equipment

1. Peptide synthesizer CS-Bio 336x or equivalent
   
2. Analytical balance
   
3. Swing bucket eppendorf centrifuge or equivalent
   
4. High performance liquid chromatograph (HPLC)
   
5. Reaction vessel (SKU: PS-G002 )
   
6. Liquid chromatograph-mass spectrometer
   

Procedure