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Recombinant Human Cytosolic beta-Glucosidase/GBA3, CF

Catalog #: 5969-GH
1 Citation Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
5969-GH-010
R&D Systems Recombinant Proteins and Enzymes
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Citations (1)
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Cytosolic beta-Glucosidase/GBA3, 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-beta -D-glucopyranoside. The specific activity is >1,500 pmol/min/μg, as measured under the described conditions.
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Cytosolic beta-Glucosidase/GBA3 protein
Thr13-Leu469 with a C-terminal 6-His tag
Accession #
Q9H227
N-terminal Sequence
Analysis
Thr13 & Pro124
Predicted Molecular Mass
53 kDa & 41 kDa
SDS-PAGE
54 kDa, reducing conditions

Product Datasheets

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5969-GH

Product Datasheet
COA
SDS

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.

5969-GH

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.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Assay Procedure

Materials
  • 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
  1. Dilute rhGBA3 to 1 ng/μL in Assay Buffer.
  2. Dilute Substrate to 800 μM in Assay Buffer.
  3. 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.
  4. Read at excitation and emission wavelengths of 365 nm and 445 nm (top read), 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 (Sigma, Catalog # M1381).

Per Well:
  • rhGBA3: 0.050 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: 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).

References
  1. Tybulewicz, V.L. et al. (1992) Nature 357:407.
  2. Matern, H. et al. (2001) J. Biol. Chem. 276:37929.
  3. de Graaf, M. et al. (2001) Biochem. J. 356:907.
  4. Hayashi, Y. et al. (2007) J. Biol. Chem. 282:30889.
Entrez Gene IDs
57733 (Human); 289687 (Rat)
Alternate Names
CBG; CBGL1; CBGL1GLUC; Cytosolic betaGlucosidase; Cytosolic beta-Glucosidase; Cytosolic beta-glucosidase-like protein 1; EC 3.2.1.21; GBA3; GLUC; glucosidase, beta, acid 3 (cytosolic); klotho-related protein; KLrP; MGC104276; MGC126878

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

  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, 2015-03-12;10(3):e0119141.
    Species: Human, Mouse
    Sample Types: Tissue Homogenates
    Applications: Enzyme Assay

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