8 m guanidine hydrochloride

We purchased the therapeutic mAb immunoglobulin (IgG) gamma 1 drugs adalimumab and rituximab from Eisai Co., Ltd. (Tokyo, Japan) and Chugai Pharmaceutical Co., Ltd. (Tokyo, Japan), respectively. In addition, we purchased Tris-HCl buffer (pH 8.0), 10% trifluoroacetic acid, 0.1% formic acid, acetonitrile 0.1% formic acid and 7 K Dialysis Casettes from Thermo Fisher Scientific (San Jose, CA, USA). We obtained 8 M guanidine hydrochloride from Sigma-Aldrich (St. Louis, MO), dithiothreitol, iodoacetamide, sodium acetate, 2-morpholinoethanesulfonic acid, 4-(2-hydroxyethyl)-1-piperazinepropanesulfonic acid and deuterium oxide from FUJIFILM Wako Pure Chemical Co., Ltd. (Tokyo, Japan), and MicroSpin G-25 columns from GE Healthcare (Chicago, IL). We purchased trypsin from Promega Co. (Madison, WI); angiotensin II from Peptide Institute, Inc. (Osaka, Japan); and ¹⁸O-water, acetonitrile (LC-MS grade), and ordinary water (LC-MS grade) from Merck (Darmstadt, HE).

12–16-week-old [18 ] female and male Cyba^nmf333 (Cyba mut) and internal Cyba^+/+ (WT) control mice on C57BL/6J background were euthanized by cervical dislocation. Intestines were excised, colon and ileum horizontally cut and rinsed carefully with PBS. Colonic (whole colon) and ileal (terminal ileum 10 cm) mucins were isolated and purified as previously published [19 (link)]. The procedure was modified by using 8M guanidine hydrochloride (Sigma) for the initial extraction procedure, and for desalting of the samples in the last step by using a Bio-Gel P-6D (Bio-Rad) column.