The synthesized linear GeXXVIIA was alkylated with 10 mM iodoacetamide (IAA) in 100 mM Tris-HCl, pH 8.7, and 2 mM EDTA at 22 °C in the dark for 1 h. The alkylated linear peptide GeXXVIIA-L was purified on a ZORBAX C18 HPLC analytical column (4.6 × 250 mm, Agilent). The digestion of alkylated GeXXVIIA-L was carried out in 100 mM Tris-HCl, pH 8.5, using 3 μg/mL Asp-N (Sigma, St Louis, MO, USA) at 37 °C for 18 h. The digested products were separated on a ZORBAX C18 HPLC analytical column (4.6 × 250 mm, Agilent).
Zorbax c18 hplc analytical column
Manufactured by Agilent Technologies
Sourced in United States
The ZORBAX C18 HPLC analytical column is a high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of organic compounds. It features a stationary phase of silica particles with a chemically bonded C18 (octadecylsilane) functionality, which provides excellent retention and separation capabilities for a variety of analytes.
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Lab products found in correlation
2 protocols using zorbax c18 hplc analytical column
1
Alkylation and Digestion of Linear Peptide
2
Oxidative Folding of Linear Peptide
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The linear peptide of GeXXVIIA, with all 5 Cys residues unprotected, was synthesized by the Chinese Peptide Company (Hangzhou, China). Oxidative folding of the purified linear peptide (20 μM) was performed at 4 °C for one week in a buffer comprising 50 mM Tris-HCl, pH 8.0, 400 mM arginine, 0.02 mM DTT, and 0.3 mM oxidized DTT. The oxidized products were purified on a ZORBAX C18 HPLC analytical column (4.6 × 250 mm, Agilent) with isocratic elution. Each monomer component was applied to a secondary HPLC purification.
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