Based on the locations of conserved Glu residues in hfgl2 sequence shown by HomoloGene and DNAStar and its nature as a serine protease, we identified a peptide NPG-12 that was named for its location at the N-terminus of membrane-bound hfgl2, its length of 12 amino acid residues (corresponding to residues 76–87), and its abundance of Glu residues [19] , [20] (link). NPG-12 scrambled peptide (NSP) comprised of the same amino acids but in different order as NPG-12 was synthesized as a control. The peptides coupling with cysteine (Cys) was synthesized by GL Biochem Ltd (China) through solid-phase chemistry (PSSM-8, Shimadzu, Japan), and high-pressure liquid chromatography (LC-20A, Shimadzu, Japan) was utilized to examine its purity. KLH (Sigma, USA) was selected as a conjugate, and Cys cross-linked with sulfo-SMCC-KLH (Pierce Chemical, USA) through a mercapto group [21] (link). Then, the peptide-KLH was used as an antigen [18] (link), [20] (link)–[23] (link).
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