Dynamics of Carbohydrate Affinities at the Cell Surface: MATERIALS AND METHODS(2)

METHODS(2)

The tetramethylbenzidine was from Merck (Darmstadt, Germany). Chlortetracycline (CTC), Toluidine Blue, Л-a-benzoyl-L-argi-nine-p-nitroanilide (4.0 mM), cartilage chondroitin 4-sulphate, heparin-al-bumin-biotin (heparin-BSA-biotin), inositol, D( + )-mannose, maltose, mannan (from Saccharomyces ceravisiae), D( + )-glucose, fucose, fucoi-dan, 1-O-methyl-a-D-glycopyranoside and bovine serum albumin (BSA) as well as all other chemicals were from Sigma (St. Louis, MO) if not indicated otherwise. buy cheap allegra

The investigations reported in this manuscript were conducted after approval of the Ethical Committee for Animal Experiments of Utrecht University and were in accordance to the International Guiding Principles for Biomedical Research Involving Animals as promulgated by the Society for the Study of Reproduction.

Semen Collection and Preparation

Semen was obtained from three fertile bulls housed at the Faculty of Veterinary Medicine, Utrecht University, The Netherlands. Freshly collected semen was diluted 10 times in saline medium (containing 137 mM NaCl, 2.5 mM KCl, 10 mM glucose, 20 mM Hepes) of 38°C and transported to the laboratory, avoiding cold-shock damage of the sperm. All buffers were iso-osmotic (285-300 mOsm) with pH 7.4 unless indicated otherwise.

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