SiaFindTM α2,6-Specific Reagent Kits (SK2601 and SK2601B) contain a recombinant protein engineered from Polyporus squamosus lectin (PSL). This highly purified affinity reagent is designed for sensitive, robust, and specific detection of Siaα2,6Gal commonly found in glycoconjugates (glycoproteins, glycolipids, and oligo- or polysaccharides). Applications include use as a primary probe in ELISA, Western blot, and immunohistochemistry. It is not recommended for flow cytometry. Each kit also includes a 5X binding buffer to ensure maximum reagent specificity and ease of use.
Each SiaFindTM 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 SiaFindTM 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.