Recombinant Human Hexosaminidase B/HEXB Protein, CF

Catalog # Availability Size / Price Qty
8907-GH-020
R&D Systems Recombinant Proteins and Enzymes
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Recombinant Human Hexosaminidase B/HEXB Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to hydrolyze 4-methylumbelliferyl-N-acetyl-beta -D-glucosaminide (4-MU-GlcNAc) The specific activity is >6,000 pmol/min/μg, as measured under the described conditions.
Source
Chinese Hamster Ovary cell line, CHO-derived human Hexosaminidase B/HEXB protein
Ala43-Met556, with C-terminal 6-His
Accession #
N-terminal Sequence
Analysis
Ala43
Predicted Molecular Mass
60 kDa
SDS-PAGE
59-70 kDa, reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

8907-GH

Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
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.

Assay Procedure

Materials
  • Assay Buffer:  0.1 M MES, pH 5.5
  • Recombinant Human Hexosaminidase B/HEXB (rhHEXB) (Catalog # 8907-GH)
  • Substrate: 4-Methylumbelliferyl-N-Acetyl-beta -D-glucosaminide (4-MU-GlcNAc) (Sigma, Catalog # M2133), 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 rhHEXB to 2 ng/μL in Assay Buffer.
  2. Dilute Substrate to 800 μM in Assay Buffer.
  3. Load into a plate 50 μL of 2 ng/μL rhHEXB, and start the reaction by adding 50 μL of 800 μM Substrate. For Substrate Blank, load 50 μL of Assay Buffer and 50 μL of 800 μM Substrate.
  4. Read plate at excitation and emission wavelengths of 365 nm and 445 nm, respectively, 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 (4-MU) (Sigma, Catalog # M1381) Per Well:
  • rhHEXB: 0.1 μg
  • Substrate: 400 μM
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Hexosaminidase B/HEXB

Beta-hexosaminidases are enzymes involved in the hydrolysis of terminal N-acetyl-D-hexosamine residues in GM2 gangliosides and globo sphingolipids in lysosomes in the brain and other tissues (1-4). These enzymes are composed of alpha and/or beta  subunits, which are coded by HEXA and HEXB genes, respectively. Different combination of alpha and beta subunits gives rise to beta-hexosaminidase isoform A, B and S (Hex A, B and S) (5), which have the composition of alpha beta, beta beta and alpha alpha, respectively. The recombinant HEXB is presumably in the B isoform and is highly active on 4-methylumbelliferyl-N-acetyl-beta -D-glucosaminide. Mutations in HEXA and HEXB genes cause lysosomal lipid storage disorders, such as Tay-Sachs disease, manifested by harmful accumulation of ganglioside GM2 in tissues and nerve cells in the brain (6-9). Also, mutation of HEXB results in Sandhoff disease manifested by the accumulation of both ganglioside GM2 and globoside in tissues and nerve cells in the brain (10, 11).

References
  1. Gilbert, F. et al.1975, Proc. Natl. Acad. Sci. USA 72:263.
  2. Myerowitz, R. et al.1985, Proc. Natl. Acad. Sci. USA 82:7830.
  3. Korneluk, R.G. et al.1986, J. Biol. Chem. 261:8407.
  4. Mark, B.L. et al.2003, J. Mol. Biol. 327:1093.
  5. Mahuran, D.J. et al.1988, J. Biol. Chem. 263:4612.
  6. Mahuran, D.J.1991, Biochim. Biophys. Acta. 1096:87.
  7. Mencarelli, S. et al.2005, FEBS Lett. 579:5501.
  8. Neufeld, E.F.1989, J. Biol. Chem. 264:10927.
  9. Ohno, K. et al.2008, Mol. Genet. Metab. 94:462.
  10. Maier T., et al. 2003 J. Mol. Biol. 328:669.
  11. Gomez-Lira M., et al. 1995 Hum. Genet. 96:417.
Entrez Gene IDs
3074 (Human); 15212 (Mouse); 294673 (Rat)
Alternate Names
beta-hexosaminidase subunit beta; Beta-N-acetylhexosaminidase subunit beta; Cervical cancer proto-oncogene 7 protein; EC 3.2.1.52; ENC-1AS; HCC-7; HEL-248; HEXB; hexosaminidase B (beta polypeptide); Hexosaminidase B; Hexosaminidase subunit B; N-acetyl-beta-glucosaminidase subunit beta

Citation for Recombinant Human Hexosaminidase B/HEXB Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. Synthesis, conformational analysis and glycosidase inhibition of bicyclic nojirimycin C-glycosides based on an octahydrofuro[3,2-b]pyridine motif
    Authors: J Désiré, Q Foucart, A Poveda, G Gourlaouen, Y Shimadate, M Kise, C Proceviat, R Ashmus, DJ Vocadlo, J Jiménez-Ba, A Kato, Y Blériot
    Carbohydrate research, 2021;511(0):108491.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay

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