Recombinant S. pneumoniae beta(1-4)-Galactosidase, CF Summary
Thr2-Leu595, with an N-terminal Met and a 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 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.
- 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
- Dilute rSp beta (1-4)-Galactosidase to 4 ng/µL in Assay Buffer.
- Dilute Substrate to 500 µM in Assay Buffer.
- 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.
- 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).
- rSp beta (1-4)-Galactosidase: 0.20 µg
- Substrate: 250 µM
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).
- Lis, H. and Sharon, N. (1993) Eur. J. Biochem. 218:1.
- Hart, G.W. (1992) Curr. Opin. Cell Biol., 4:1017.3.
- Dwek, R.A. (1995) Biochem. Soc. Trans. 23:1.
- Wormald, M.R. and Dwek, R.A. (1999) Structure 7:R155.
- Hood, J.M. et al. (1977) Proc. Natl. Acad. Sci. USA 75:113.
- Glasgow, L. et al. (1977) J. Biol. Chem. 252:8615.
No product specific FAQs exist for this product, however you mayView all Proteins and Enzyme FAQs
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/¥75 Yuan/¥1250 Yen for a review with an image
$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image