Recombinant X. campestris beta(1-3)-Galactosidase, CF

R&D Systems | Catalog # 5704-GH

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Key Product Details

  • R&D Systems E. coli-derived Recombinant X. campestris beta(1-3)-Galactosidase (5704-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

E. coli

Accession Number

NP_638243

Applications

Enzyme Activity
View all beta (1-3)-Galactosidase Proteins and Enzymes »

Product Specifications

Source

E. coli-derived x. campestris beta (1-3)-Galactosidase protein
Ser25-Glu613 with an N-terminal Met and 6-His tag
Accession # NP_638243

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

67 kDa

SDS-PAGE

63 kDa, reducing conditions

Activity

Measured by its ability to cleave a fluorogenic substrate, 4-Methylumbelliferyl-beta -D-galactopyranoside.
The specific activity is >14,000 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

5704-GH
Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Shipping The product is shipped with dry ice or equivalent. 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: beta (1-3)-Galactosidase

The majority of secreted and membrane proteins are glycosylated (1, 2). Proper glycosylation might be critical for protein folding and biological functions (3, 4). Galactoside is an essential sugar commonly found on various glycan conjugates and galactosidases are among the earliest enzymes to be studied (5). beta 1‑3 Galactosidase from Xanthomanas capestris is a useful tool for removing beta 1-3 linked galactosides from the non-reducing terminus of glycoconjugates (6, 7).

References

  1. Lis, H. and Sharon, N. (1993) Eur. J. Biochem. 218:1.
  2. Hart, G.W. (1992) Curr. Opin. Cell Biol. 4:1017.
  3. Dwek, R.A. (1995) Biochem. Soc. Trans. 23:1.
  4. Wormald, M.R. and Dwek, R.A. (1999) Structure 7:R155.
  5. Hood, J.M. et al. (1977) Proc. Natl. Acad. Sci. USA 75:113.
  6. Taron, C. et al. (1995) Glycobiology 5:603.
  7. Glasgow, L. et al. (1977) J. Biol. Chem. 252:8615.

Alternate Names

beta (13)Galactosidase

UniProt

NP_638243

Additional beta (1-3)-Galactosidase Products

Product Documents for Recombinant X. campestris beta(1-3)-Galactosidase, 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 X. campestris beta(1-3)-Galactosidase, CF

For research use only

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Protocols

View specific protocols for Recombinant X. campestris beta(1-3)-Galactosidase, CF (5704-GH):

Materials
  • Assay Buffer: 0.1 M MES, pH 5.5
  • Recombinant X. campestris beta (1‑3)-Galactosidase (rXc beta -Galactosidase) (Catalog # 5704-GH)
  • Substrate: 4-methylumbelliferyl-beta -D-galactopyranoside (Sigma, Catalog # M1633), 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 rXc beta -Galactosidase to 1 ng/µL in Assay Buffer.
  2. Dilute Substrate to 400 µM in Assay Buffer.
  3. Load into plate 50 µL of 1 ng/µL rXc beta -Galactosidase, and start the reaction by adding 50 µL of 400 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL 400 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 (pmoles/min/µg) =

 Adjusted Vmax* (RFU/min) x Conversion Factor** (pmole/RFU)
amount of enzyme (µg)

     *Adjusted for Substrate Blank

     **Derived using calibration standard 4-methylumbelliferone (Sigma, Catalog # M1381).

Per Well:
  • rXc beta -Galactosidase: 0.050 µg
  • Substrate: 200 µM

FAQs

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