Recombinant Human Hexosaminidase A/HEXA Protein, CF SummaryLearn more about Fluorescent Glycan Labeling and Detection
Met1-Thr529, with a C-terminal 6-His tag
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.
|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.
- Assay Buffer: 100 mM Sodium Citrate, 250 mM NaCl, pH 4.5
- Recombinant Human Hexosaminidase A/HEXA (rhHEXA) (Catalog # 6237-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 rhHEXA to 5 ng/μL in Assay Buffer.
- Dilute Substrate to 800 μM in Assay Buffer.
- Load into a plate 50 μL of 5 ng/μL rhHEXA, and start the reaction by adding 50 μL of 800 μM Substrate. For Substrate Blanks, 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).
- rhHEXA: 0.250 μg
- Substrate: 400 μM
Background: Hexosaminidase A/HEXA
beta -hexosaminidases are enzymes involved in the hydrolysis of terminal N-acetyl-D-hexosamine residues in GM2 gangliosides and globo-sphingolipids in lysosomes
(1‑4). The enzymes are composed of two alpha and/or beta subunits, which are coded by HEXA and HEXB genes, respectively. Different association of the alpha and beta subunits gives rise to beta ‑hexosaminidase isoforms A, B and S (Hex A, B and S) (5), which have the composition of alpha beta, beta beta and alpha alpha, respectively. Our recombinant HEXA is presumably isoform Hex S, because only alpha subunit was expressed. Hex S is suggested to releases non‑reducing end N-acetylgalactosamine residues from dermatan sulfate, chondroitin sulfate and sulfated glycolipid SM2 (6). Recombinant HEXA is also highly active on 4-methylumbelliferyl-N-acetyl-beta -D-glucosaminide (6). Mutations in HEXA and HEXB genes cause lysosomal lipid storage disorders. Specifically, mutations of HEXA cause Tay-Sachs disease, manifested by the harmful accumulation of ganglioside GM2 in tissues and nerve cells in the brain (7‑10). Children with this disease usually die by age 4.
- 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.
- Hepbildikler, S.T. et al. (2002) J. Biol. Chem. 277:2562.
- 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.
Citations for Recombinant Human Hexosaminidase A/HEXA 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.
Citations: Showing 1 - 4
Filter your results:
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.
Sample Types: Protein
Bi-functional IgG-lysosomal enzyme fusion proteins for brain drug delivery
Authors: RJ Boado, JZ Lu, EK Hui, H Lin, WM Pardridge
Sci Rep, 2019;9(1):18632.
Sample Types: Protein
Identification and characterization of the Onchocerca volvulus Excretory Secretory Product Ov28CRP, a putative GM2 activator protein
Authors: FN Njume, SM Ghogomu, RA Shey, LOT Gainkam, P Poelvoorde, P Humblet, J Kamgno, A Robert, L Mutesa, C Lelubre, E Edelweiss, A Poterszman, S Anheuser, L Vanhamme, J Souopgui
PLoS Negl Trop Dis, 2019;13(7):e0007591.
Species: Nematode - Onchocerca volvulus
Sample Types: Recombinant Protein
Human neutrophils secrete bioactive paucimannosidic proteins from azurophilic granules into pathogen-infected sputum.
Authors: Thaysen-Andersen M, Venkatakrishnan V, Loke I, Laurini C, Diestel S, Parker B, Packer N
J Biol Chem, 2015;290(14):8789-802.
Sample Types: Protein
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