N-GlyFind™ Core-Specific Reagent Kits (NK0101 and NK0101B) contain a recombinant protein engineered from Mus musculus Fbs1 (Fbxo2), a component of the E3 ubiquitin ligase complex. This highly purified affinity reagent is designed for sensitive, robust, and specific detection of the N-glycan core structure (Man3GlcNAc2-Asn) commonly found in glycoproteins. Applications include use as a primary probe in ELISA, Western blot, and other assays. Each kit also includes 5X Tris Buffered Saline (BA0104) as a binding buffer to ensure maximum reagent specificity and ease of use.
Each N-GlyFind™ Core-Specific Reagent has a molecular mass of about 26 kD and works without bivalent metal ions. It is 8xHis-tagged at its N-terminus, 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 N-GlyFind™ 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 10 mg/mL solution. Concentration is determined by spectrophotometry using E1% 26.4. 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.