Recombinant Human MAN2B1 Protein, CF

R&D Systems | Catalog # 9826-GH

R&D Systems
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Key Product Details

  • R&D Systems HEK293-derived Recombinant Human MAN2B1 Protein (9826-GH)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

HEK293

Accession Number

Applications

Enzyme Activity
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Product Specifications

Source

Human embryonic kidney cell, HEK293-derived human MAN2B1 protein
Gly50-Gly1011, with a C-terminal 6-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Gly50

Predicted Molecular Mass

110 kDa

SDS-PAGE

109-127 kDa, reducing conditions

Activity

Measured by its ability to hydrolyze 4-methylumbelliferyl-alpha -D-mannopyranoside.
The specific activity is >1,500 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

9826-GH
Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: MAN2B1

The lysosomal catabolism of N-glycans on mammalian glycoproteins occurs through the sequential exoglycosidase digestion of oligosaccharides from the non‑reducing terminus down to the carbohydrate-peptide core region (1). Included among the hydrolytic activities responsible for this oligosaccharide degradation is a broad specificity exo-alpha -mannosidase MAN2B1 that catalyzes the hydrolysis of alpha 1,2-, alpha 1,3-, and alpha 1,6-mannoside linkages present in complex, hybrid, and high mannose Asn-linked glycans (2, 3, 4). Defects in MAN2B1 result in lysosomal alpha -mannosidosis that shows symptom of mental retardation, recurrent infections, impaired hearing and Hurler-like skeletal changes (5, 6).

Long Name

Mannosidase alpha, Class 2B, Member 1

Alternate Names

LAMAN, MANB, Mannosidase Alpha-B

Entrez Gene IDs

4125 (Human); 17159 (Mouse); 361378 (Rat)

Gene Symbol

MAN2B1

UniProt

Additional MAN2B1 Products

Product Documents for Recombinant Human MAN2B1 Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Manufacturing Specifications

  1. Aronson, N.N. Jr. and Kuranda, M.J. (1989) FASEB J. 3:2615.
  2. Liao, Y.F. et al. (1996) J. Biol. Chem. 271:28348.
  3. Gotoda, Y. et al. (1989) Am. J. Hum. Genet. 63:1015.
  4. Nebes, V.L. and Schmidt, M.C. (1994) Biochem. Biophys. Res. Commun. 200:239.
  5. Nilssen, O, et al. (1997) Hum. Mol. Genet. 6:717.
  6. Yu, P. et al. (2015) Genet. Med. 17:971.

Product Specific Notices for Recombinant Human MAN2B1 Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant Human MAN2B1 Protein, CF (9826-GH):

Materials
  • Assay Buffer: 50 mM Sodium Acetate, pH 4.5
  • Recombinant Human MAN2B1 (rhMAN2B1) (Catalog # 9826-GH)
  • Substrate: 4-Methylumbelliferyl-alpha -D-mannopyranoside (Sigma, Catalog # M3657), 50 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhMAN2B1 to 2 ng/μL in Assay Buffer.
  2. Dilute Substrate to 400 µM in Assay Buffer.
  3. Load 50 μL of 2 ng/μL rhMAN2B1 into the plate, and start the reaction by adding 50 μL of 400 µM Substrate. Include a Substrate Blank containing 50 μL of Assay Buffer and 50 μL of 400 µM Substrate.
  4. Read at excitation and emission wavelengths of 365 nm and 445 nm, respectively, (top read) in kinetic mode for 5 minutes.
  5. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

*Adjusted for Substrate Blank.
**Derived using calibration standard 4-Methylumbelliferone (Sigma, Catalog # M1381).

Per Well:
  • rhMAN2B1: 0.1 µg
  • Substrate: 200 µM

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