Envicarb 100 mg

Eluents with increasing buffer concentration were compared. The mobile phase was composed of ethyl lactate (35%, (v/v)) mixed with a phosphate buffer (60 to 200 mM, pH 8) containing 10 mM sodium sulfite. In triplicate, Supelco EnviCarb 100 mg cartridges were conditioned with 2.4 mL ethanol, then reduced with 2.4 mL of 60 mM metabisulfite. Cartridges were loaded with 24 mL of BW, washed with 2.4 mL of deionized water, then eluted with 1.2 mL of mobile phase. Marennine concentration was determined by UV–Vis, by comparing the absorbance of each sample at the λ_max in the visible range, with a cuvette of 1 cm path length.

Aliquots (150 μg) of mouse monoclonal anti-MBP antibody (New England Biolabs), human or rabbit polyclonal IgG (Sigma-Aldrich) were incubated with Rapid PNGase F (New England Biolabs) in a total volume of 50 μL of Rapid PNGase F reaction buffer for 10 min at 50 °C. For use in lectin ELISA experiments, the deglycosylated antibodies were dialyzed against 20 mM sodium phosphate, pH 7.4, 150 mM NaCl using 10,000 Da MWCO dialysis membranes. For glycan labeling, the liberated N-glycans were cleaned and desalted using SPE Supelclean columns ENVI-18 100 mg and ENVI-CARB 100 mg (Supelco).