In competitive binding assays, 10 mm free synthetic sialic acid (Sigma) was added 30 min before the endocytic agent and left during the assay. These were further purified by size-exclusion chromatography with elution in 50 mm KPi, 200 mm NaCl, pH 7.8, dialyzed against assay buffer (20 mm Tris, 150 mm NaCl, 2 mm tris(2-carboxyethyl)phosphine, pH 7.5), and finally concentrated to 0. If you have any queries relating to where by and how to use manufacturer of sialic acid powder as Raw Material for food, you can get hold of us at our own webpage. 5-1 mm for storage at 4 °C. All 1D NMR experiments were performed using a Bruker Advance I 500-MHz spectrometer with a 5-mm PATXI 1H/D-13C/15N Z-GRD probe at 293 K.