Supplementary Information

Each SiaFind^TM α2,6-Specific Reagent has a molecular mass of about 34 kD and works with or without bivalent metal ions. They are N-terminal 8xHis-tagged proteins, and an anti-polyhistidine antibody or, in the case of the biotinylated version, a streptavidin conjugate can be used for detection. The 8xHis tag may be removed by FasTEV (Cat #GE0501), a TEV protease with enhanced stability and catalytic activity.

Form and Storage

The SiaFind^TM reagents are supplied lyophilized in a storage buffer (50 mM EPPS, 200 mM NaCl, pH 7.5) and should be reconstituted in 100 μL molecular water grade to yield a 1 mg/mL solution. Concentration is determined by spectrophotometry using E1% 22.0. Once reconstituted, store at 4°C for up to 5 days or -20°C for up to 6 months. Aliquoting is recommended to avoid repeated freeze-thaw cycles.

All 5X buffers should be diluted to 1X with ultrapure water. For instance, to make 250 mL, add 50 mL of any 5X buffer to 200 mL water and mix by inversion. All buffers may be stored at 4 to 25°C.

Western Blot