Recombinant S. pneumoniae beta(1-4)-Galactosidase, CF

R&D Systems | Catalog # 5549-GH

R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems E. coli-derived Recombinant S. pneumoniae beta(1-4)-Galactosidase (5549-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

YP_815516

Applications

Enzyme Activity
View all beta (1-4)-Galactosidase Proteins and Enzymes »

Product Specifications

Source

E. coli-derived s. pneumoniae beta (1-4)-Galactosidase protein
Thr2-Leu595, with an N-terminal Met and a 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

Met

Predicted Molecular Mass

70 kDa

SDS-PAGE

62 kDa, reducing conditions

Activity

Measured by its ability to cleave a fluorogenic substrate, 4-Methylumbelliferyl-beta -D-galactopyranoside.
The specific activity is >600 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

5549-GH
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.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Background: beta (1-4)-Galactosidase

Majority of secreted and membrane proteins are glycosylated (1, 2). Proper glycosylation could be critical for protein folding and biological functions (3, 4). Galactosides are commonly found on various glycan conjugates and galactosidases are tools for removing these sugar residues (5). beta 1-4 Galactosidase from Streptococcus pneumoniae is especially efficient in removing beta 1-4 linked non-reducing galactosides from Gal-beta 1-4-GlcNAc or Gal-beta 1-4-GalNAc linkage (6).

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.
  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. Glasgow, L. et al. (1977) J. Biol. Chem. 252:8615.

Alternate Names

beta (14)Galactosidase, beta-Galactosidase-3

UniProt

YP_815516

Additional beta (1-4)-Galactosidase Products

Product Documents for Recombinant S. pneumoniae beta(1-4)-Galactosidase, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Recombinant S. pneumoniae beta(1-4)-Galactosidase, CF

For research use only

Citations for Recombinant S. pneumoniae beta(1-4)-Galactosidase, CF

Customer Reviews for Recombinant S. pneumoniae beta(1-4)-Galactosidase, CF

There are currently no reviews for this product. Be the first to review Recombinant S. pneumoniae beta(1-4)-Galactosidase, CF and earn rewards!

Have you used Recombinant S. pneumoniae beta(1-4)-Galactosidase, CF?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

Protocols

View specific protocols for Recombinant S. pneumoniae beta(1-4)-Galactosidase, CF (5549-GH):

Materials
  • Assay Buffer: 50 mM Sodium Phosphate, pH 6.0
  • Recombinant S. pneumoniae beta (1‑4)-Galactosidase (rSp beta (1-4)-Galactosidase) (Catalog # 5549-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 rSp beta (1-4)-Galactosidase to 4 ng/µL in Assay Buffer.
  2. Dilute Substrate to 500 µM in Assay Buffer.
  3. Load into plate 50 µL of 4 ng/µL rSp beta (1-4)-Galactosidase and start the reaction by adding 50 µL of 500 µM Substrate. Include a Substrate Control containing 50 µL Assay Buffer, 50 µL of 500 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:
  • rSp beta (1-4)-Galactosidase: 0.20 µg
  • Substrate: 250 µM

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes