Recombinant Human Cytosolic beta-Glucosidase/GBA3, CF

R&D Systems | Catalog # 5969-GH

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Key Product Details

  • R&D Systems Sf 21 (baculovirus)-derived Recombinant Human Cytosolic beta-Glucosidase/GBA3 (5969-GH)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

Sf 21 (baculovirus)

Accession Number

Q9H227

Applications

Enzyme Activity
View all Cytosolic beta-Glucosidase/GBA3 Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Cytosolic beta-Glucosidase/GBA3 protein
Thr13-Leu469 with a C-terminal 6-His tag

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.

N-terminal Sequence Analysis

Thr13 & Pro124

Predicted Molecular Mass

53 kDa & 41 kDa

SDS-PAGE

54 kDa, reducing conditions

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.

Formulation, Preparation, and Storage

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.

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.

Alternate Names

CBGL1, Cytosolic betaGlucosidase, GBA3, GLUC, KLrP

Entrez Gene IDs

57733 (Human); 289687 (Rat)

Gene Symbol

GBA3

UniProt

Q9H227 (Human)

Additional Cytosolic beta-Glucosidase/GBA3 Products

Product Documents for Recombinant Human Cytosolic beta-Glucosidase/GBA3, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human Cytosolic beta-Glucosidase/GBA3, CF

For research use only

Citations for Recombinant Human Cytosolic beta-Glucosidase/GBA3, CF

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Protocols

View specific protocols for Recombinant Human Cytosolic beta-Glucosidase/GBA3, CF (5969-GH):

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

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