Sialidase Cp α-(2-3,6) - TEST
α(2-3,6) Sialidase Cp cleaves all non-reducing terminal non-branched α(2-3) and α(2-6) sialic acid residues from complex carbohydrates and glycoproteins. There is no detectable activity on α(2-8) or α(2-9) linkages or on branched α(2-3) or α(2-6) linkages. The relative cleavage rates for different linkages are: α(2-3) > α(2-6).
α(2-3,6) Sialidase Cp will not cleave branched sialic acids (linked to an internal residue). Use α(2-3,6,8,9) Sialidase Au (E-S001) for α(2-8) or branched sialic acids. To cleave only non-reducing terminal α(2-3) unbranched sialic acid residues, use α(2-3) Sialidase Sp (E-S007).
α(2-3,6) Sialidase Cp is isolated from a clone of Clostridium perfringens. The enzyme has been extensively characterized using oligosaccharide standards.
Alternate Names: Neuraminidase, NANase, N-acetylneuraminate glycohydrolase, Exo-α-sialidase
Source: Recombinant from Clostridium perfringens in E. coli.
Sialidase Cp in 20 mM Tris-HCl, 25 mM NaCl, pH 7.5
5x Reaction Buffer 250 mM sodium phosphate, pH 6.0
Specific Activity: >250 U/mg
Activity: >15 U/mL
Molecular weight: 41,000 daltons
pH range: 50 mM sodium phosphate (pH 6.0) provides the optimal buffer for enzyme activity with 3'-siayllactose, a standard substrate. If glycosidase treatment is performed at suboptimal pH because of glycoprotein solubility or activity requirements, expect some diminution in enzyme activity.
1. Add up to 100 µg of glycoprotein or 1 nmol of oligosaccharide to tube.
2. Add water to 14 µL
3. Add 4 µL 5X Reaction Buffer.
4. Add 2 µL α(2-3,6) Sialidase Cp.
5. Incubate at 37°C for 1 hour.
Desialylation may be monitored by SDS-PAGE if the size differential between native and desialylated protein is sufficient for detection.
Specificity: Cleaves all nonreducing terminal non-branched α(2-3) and α(2-6) sialic acid residues from complex carbohydrates and glycoproteins.
Relative cleavage rates for different linkages are: (2-3) > (2-6).
Specific Activity Assay: Defined as the amount of enzyme required to produce 1 µmole of methylumbelliferone in 1 minute at 37°C, pH 5.0 from MU-NANA [2'-(4-methylumbelliferyl)-α-D-N-acetylneuraminic acid].
Storage: Store enzyme at 4°C.