Recombinant Human Cytosolic beta-Glucosidase/GBA3, CF Summary
Thr13-Leu469 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, NaCl, DTT 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 Buffer: 0.1 M MES, pH 6.5
- Recombinant Human Cytosolic beta ‑Glucosidase/GBA3 (rhGBA3) (Catalog # 5969-GH)
- Substrate: 4-methylumbelliferyl-beta -D-glucopyranoside (Sigma, Catalog # M3633), 10 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhGBA3 to 1 ng/μL in Assay Buffer.
- Dilute Substrate to 800 μM in Assay Buffer.
- Load in a plate 50 μL of 1 ng/μL rhGBA3, and start the reaction by adding 50 μL of 800 μM Substrate. Include a control containing 50 μL of Assay Buffer and 50 μL of 800 μM Substrate.
- Read at excitation and emission wavelengths of 365 nm and 445 nm (top read), 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 (Sigma, Catalog # M1381).
- rhGBA3: 0.050 μg
- Substrate: 400 μM
Background: Cytosolic beta-Glucosidase/GBA3
There are three beta ‑glucosidases (GBA) in human genome. GBA1 endodes a lysosomal membrane protein that cleaves the beta ‑glucosidic linkage of glucosylceramide (1). GBA2 encodes a microsomal beta ‑glucosidase that catalyzes the hydrolysis of bile acid 3-O-glucosides (2). GBA3 is a cytosolic beta ‑Glucosidase and is predominantly expressed in liver. GBA3 efficiently hydrolyzes beta ‑D‑glucoside and beta ‑D‑galactoside, but not any known physiological beta ‑glycoside, suggesting that it may be involved in detoxification of plant glycosides (3). GBA3 also has significant neutral glycosylceramidase activity, suggesting that it may be involved in a nonlysosomal catabolic pathway of glucosylceramide metabolism (4). At the protein level, GBA3 shows significant homology (>40%) with Klotho protein that is known for its association with aging process (3, 4).
- Tybulewicz, V.L. et al. (1992) Nature 357:407.
- Matern, H. et al. (2001) J. Biol. Chem. 276:37929.
- de Graaf, M. et al. (2001) Biochem. J. 356:907.
- Hayashi, Y. et al. (2007) J. Biol. Chem. 282:30889.
Citation for Recombinant Human Cytosolic beta-Glucosidase/GBA3, 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
Tool compounds robustly increase turnover of an artificial substrate by glucocerebrosidase in human brain lysates.
Authors: Berger, Zdenek, Perkins, Sarah, Ambroise, Claude, Oborski, Christin, Calabrese, Matthew, Noell, Stephen, Riddell, David, Hirst, Warren D
PLoS ONE, 2015;10(3):e0119141.
Sample Types: Tissue Homogenates
Applications: Enzyme Assay
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