Recombinant Human Hexosaminidase B/HEXB Protein, CF
Recombinant Human Hexosaminidase B/HEXB Protein, CF Summary
Product Specifications
Ala43-Met556, with C-terminal 6-His
Analysis
Product Datasheets
Carrier Free
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.
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.
|
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
- Dilute rhHEXB to 2 ng/μL in Assay Buffer.
- Dilute Substrate to 800 μM in Assay Buffer.
- 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.
- Read plate at excitation and emission wavelengths of 365 nm and 445 nm, respectively, in kinetic mode for 5 minutes.
- 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
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).
- Gilbert, F. et al.1975, Proc. Natl. Acad. Sci. USA 72:263.
- Myerowitz, R. et al.1985, Proc. Natl. Acad. Sci. USA 82:7830.
- Korneluk, R.G. et al.1986, J. Biol. Chem. 261:8407.
- Mark, B.L. et al.2003, J. Mol. Biol. 327:1093.
- Mahuran, D.J. et al.1988, J. Biol. Chem. 263:4612.
- Mahuran, D.J.1991, Biochim. Biophys. Acta. 1096:87.
- Mencarelli, S. et al.2005, FEBS Lett. 579:5501.
- Neufeld, E.F.1989, J. Biol. Chem. 264:10927.
- Ohno, K. et al.2008, Mol. Genet. Metab. 94:462.
- Maier T., et al. 2003 J. Mol. Biol. 328:669.
- Gomez-Lira M., et al. 1995 Hum. Genet. 96:417.
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
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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, 2021-12-20;511(0):108491.
Species: Human
Sample Types: Protein
Applications: Bioassay
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