Recombinant E. coli alpha-Galactosidase/a-Gal Protein, CF

Catalog # Availability Size / Price Qty
6415-GH-020
Product Details
FAQs
Supplemental Products
Reviews

Recombinant E. coli alpha-Galactosidase/a-Gal Protein, CF Summary

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to hydrolyze 4-methylumbelliferyl-alpha -D-galactopyranoside. The specific activity is >3,000 pmol/min/μg, as measured under the described conditions.
Source
E. coli-derived e. coli alpha-Galactosidase/a-Gal protein
Val2-Val708, with an N-terminal Met and 6-His tag
Accession #
N-terminal Sequence
Analysis
Met  
Predicted Molecular Mass
82 kDa
SDS-PAGE
70-75 kDa, reducing conditions

Product Datasheets

6415-GH

Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl 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, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Assay Procedure

Materials
  • Assay Buffer: 100 mM MES, pH 6.5
  • Recombinant E. coli alpha ‑Galactosidase/ alpha ‑Gal (rE. coli alpha -Gal) (Catalog # 6415-GH)
  • Substrate: 4-Methylumbelliferyl-alpha -D-galactopyranoside (Sigma, Catalog # M7633), 6.7 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 rE. coli alpha -Gal to 0.5 ng/μL in Assay Buffer.
  2. Dilute Substrate to 800 μM in Assay Buffer.
  3. Load into a plate 50 μL of 0.5 ng/μL rE. coli alpha -Gal, and start the reaction by adding 50 μL of 800 μM Substrate. For Substrate Blanks, load 50 μL of Assay Buffer and 50 μL of 800 μM Substrate.
  4. Read plate at excitation and emission wavelengths of 365 nm and 445 nm, 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 (4-MU) (Sigma, Catalog # M1381).

Per Well:
  • rE. coli alpha -Gal: 0.025 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.

=
÷

Background: alpha-Galactosidase/a-Gal

alpha -Galactoside is prevalent in animals and plants. The alpha -Galactosidase ( alpha -Gal) from Escherichia coli is a useful tool for the removal of alpha 1,3-linked and alpha 1,6‑linked galactosides from the non-reducing terminus of complex carbohydrates and glycoproteins (1). alpha -Gal efficiently hydrolyzes raffinose, an alpha -D-galactosylsucrose, to D‑galactose and sucrose. The enzyme can also hydrolyze other alpha -Galactosides such as melibiose, stachyose, verbascose, and galactinol. The enzyme does not cleave beta -linked galactose, such as lactose.

References
  1. Schmid, K. and Schmitt, R. (1976) Eur. J. Biochem. 67:95.
  2. Spangenberg, P. et al. (2000) Carbohydr. Res. 329:65.
Entrez Gene IDs
5589615 (E. coli)
Alternate Names
alphaGalactosidase; alpha-Galactosidase; rafA

FAQs

No product specific FAQs exist for this product, however you may

View all Proteins and Enzyme FAQs

Reviews for Recombinant E. coli alpha-Galactosidase/a-Gal Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant E. coli alpha-Galactosidase/a-Gal Protein, CF and earn rewards!

Have you used Recombinant E. coli alpha-Galactosidase/a-Gal Protein, 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

Submit a Review