alpha-D-mannosidase, p-nitrophenyl-alpha-mannosidase, alpha-D-mannopyranosidase, alpha mannosidase, exo-alpha-mannosidase
Alpha Mannosidase from Jack Bean cleaves α(1-2,3,6)-linked mannose.
This enzyme is often used in conjunction with Core Alpha-(1-6) Mannosidase (E-AM02) if a noncleavable core α(1-6) mannose is present on the substrate.
Source Jack Bean
EC 3.2.1.24
Contents
Alpha-(1-2,3,6) Mannosidase in 150 mM sodium phosphate, 0.1 mM ZnCl2 pH 7.5. (pH 7.5).
Included with 20 µL and 60 µL pack sizes:
200 µl 5x Reaction Buffer 5.0 (250 mM sodium phosphate, pH 5.0).
Specific Activity >3 U/mg
Activity >10 U/ml
Molecular weight two polypeptides of 64,000 and 44,000 daltons
Suggested usage
1. Add up to 1 nmol of oligosaccharide.
2. Add deionized water to 15 µl.
3. Add 4 µl 5x Reaction Buffer 5.0.
4. Add 1 µL of enzyme
5. Incubate ten minutes at 37˚C.
Specificty All Alpha-(1-2,3,6)-linked mannose
Specific Activity Assay
One unit of Alpha Mannosidase is defined as the amount of enzyme required to hydrolyze 1 µmole of p-nitrophenyl-alpha-p-mannoside to p-nitrophenol in 1 minute at pH 5.0 and 37˚C.
Storage Store enzyme at 4˚C.
Purity Each lot of α(1-2,3,6) mannosidase is tested for contaminating substances by incubating the enzyme for 24 hours at 37°C with substrates indicated in the table below. No detectable activity is evident for any of these potential contaminants. The detection limit of this assay is 5 μU/mL (IUB).
For the protease assay, 10 μg of denatured BSA is incubated for 24 hours with 2 μl of enzyme. Analysis of the BSA band after SDS-PAGE should show no evidence of degradation.
Stability Stable at least 12 months when stored properly. Several days exposure to ambient temperatures will not reduce activity.
Alpha Mannosidase References
Snaith, S.M., and G.A. Levvy. Purification and Properties of α-D- Mannosidase from Jack-Bean meal. Biochem. J. 110: 633-670 (1968)Ganesh Kumar, B.S., Pohlrntz, G., Shculte, M., Mormann, M. Siva Kumar, N. Jack bean α-mannosidase: amino acid sequencing and N-glycosylation analysis of a valuable glycomics tool. Glycobiology 24(3):252-61 (2014)
