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Recombinant Human Hexosaminidase B/HEXB Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

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Recombinant Human Hexosaminidase B/HEXB Protein, CF Summary

Details of Functionality
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
Ala43
Protein/Peptide Type
Recombinant Enzymes
Gene
HEXB
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Dilutions
  • Enzyme Activity
Theoretical MW
60 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
59-70 kDa, reducing conditions
Publications
Read Publication using
8907-GH in the following applications:

Packaging, Storage & Formulations

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.
Buffer
Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Assay Procedure
  • 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

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Hexosaminidase B/HEXB Protein, CF

  • 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

Background

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).
  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.

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Publications for HEXB (8907-GH)(1)

We have publications tested in 1 confirmed species: Human.

We have publications tested in 1 application: Bioassay.


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Bioinformatics

Gene Symbol HEXB
Uniprot