MIITX1-Mg1a was prepared at a 100-μmol scale using Fmoc (9-fluorenyl methoxycarbonyl) SPPS on Protide Rink Amide resin (CEM Corporation). The peptide was elongated on a CEM Liberty Blue automated microwave peptide synthesizer [4-min coupling cycle: 2-min coupling (90°C), 1-min deprotection (90°C), 1-min associated washes and liquid handling] using a fivefold excess of Fmoc amino acid, ethyl cyanohydroxyiminoacetate (Oxyma), and diisopropylcarbodiimide. Fifty micromoles of peptide was cleaved from the resin and deprotected with a TFA-based cleavage solution [TFA/triisopropylsilane/H2O (95:2.5:2.5)] for 2 hours at room temperature and then worked up with diethyl ether and lyophilized. MIITX1-Mg1a was purified from the crude material [to ≥95% with a yield of 11.2 mg (7.1%)] using a 20 to 40% solvent B (99.9% ACN and 0.1% TFA) gradient over 40 min (15 ml/min−1) on a Waters Sunfire C18 OBD preparative column (pore size, 5 μm; 19 mm × 150 mm). The identity of the peptide was confirmed by MS using a Shimadzu UPLC-ESI-MS 2020 instrument (Theoretical [M+2H]2+, 1574.4; observed [M+2H]2+, 1574.4), and purity was determined on a Waters Acquity UPLC system (Acquity C18 BEH; pore size, 1.7 μm; column, 2.1 mm × 50 mm) at 0 to 70% solvent B (99.9% ACN and 0.1% TFA) over 5 min at 0.6 ml min−1.

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.