Ludger Alpha-(1-3,6)-galactosidase enzyme structure

    α(1-3,6)-galactosidase


    E-AG02    *formerly E-AG01

    α Galactosidase from E. coli cleaves α(1-3) and α(1-6)-linked, non-reducing terminal galactose from complex carbohydrates and glycoproteins.

    recombinant from E. coli in E. coli

     View product documentation: Specsheet / CofA / MSDS

    • Part Number
    • E-AG02
    • Amount of Enzyme
    • 0.4 U / 60 µL








    Kit includes enzyme plus reaction buffer.
    Sufficient for up to 60 reactions.






    Incorporate glycan standards as process controls for reliable digestion


    Ludger Exoglycosidase Alpha Gal data

    2-AB labelled Galα(1-3)Gal standard (CAB-ALPHA-GAL-01) is recommended as a process positive control in order to assess the effectiveness of a digestion.

    Shift in retention time of the standard, corresponding to digestion of α, but not β-Gal bond indicates that the enzyme is working as expected.
















    Product Specifications:


    α Galactosidase from E. coli cleaves α(1-3) and α(1-6)-linked, non-reducing terminal galactose from complex carbohydrates and glycoproteins. There is no activity on α(1-4) linked galactose. It is particularly efficient for removing α-linked galactose under conditions where the pH must be neutral or above, for example, with living cells.

    For cleaving β Galactosidase we recommend β(1-4) Galactosidase (E-BG07), recombinant from Streptococcus pneumoniae or β(1,3,4,6) Galactosidase (E-BG02), purified from bovine testes.

    Source: Recombinant from E. coli in E. coli

    EC: 3.2.1.22

    Alternate Names: α-D-galactoside galactohydrolase, melibiase

    Contents:
    Ludger Alpha-(1-3,6)-Galactosidase - Kit Contents
    α Galactosidase enzyme in 50 mM sodium phosphate, pH 7.5
    5x Reaction Buffer – 250 mM sodium phosphate, pH 6.5

    Specific Activity: >30 U/mg
    Activity: 80 U/mL

    Molecular weight: ~80,000 daltons

    Suggested Usage:
    1. Add up to 100 µg of glycoprotein or 1 nmol of oligosaccharide to tube.
    2. Add water to 13 µL
    3. Add 4 µL 5X Reaction Buffer.
    4. Add 2 µL α(1-3,6) galactosidase.
    5. Incubate at 37°C for 1 hour.

    NOTE: Longer incubations are necessary if fucose is present on the penultimate sugar.

    Specificity: Cleaves α(1-3) and α(1-6)-linked, non-reducing terminal galactose from complex carbohydrates and glycoproteins.

    Specific Activity: One unit of α(1-3,6)-Galactosidase is defined as the amount of enzyme required to produce 1 µmole of p-nitrophenol (pNP) in 1 minute at 25°C pH 6.5 from p-nitrophenyl-alpha-D-galactopyranoside.

    Storage: Store enzyme at 4°C.

    Purity: Each lot of α Galactosidase is tested for contaminating protease as follows; 10 µg of denatured BSA is incubated for 24 hours with 2 µL of enzyme. SDS-PAGE analysis of the treated BSA shows no evidence of degradation.

    The production host strain has been extensively tested and does not produce any detectable glycosidases.

    Stability: Stable at least 12 months when stored properly. Several days exposure to ambient temperatures will not reduce activity.





Companion Products:

Process controls - View our N-glycan Nomenclature Table


Glycan clean-up - removal of enzymes from glycan mixtures following exoglycosidase treatment

  • LC-EXO-A6
  • LC-EXO-96
  • LudgerClean Post-Exoglycosidase Clean-up Spin Columns (6 samples)
  • LudgerClean Post-Exoglycosidase Clean-up Plate (96 samples)

Labelling and derivatisation of released glycans - View our Glycan Labelling Summary Table

  • LT-KAB-A2
  • LT-KAB-VP24
  • LT-KAB-VP96
  • LT-KPROC-24
  • LT-KPROC-96
  • LT-KPROC-VP24
  • LT-KAA-A2
  • LT-KAA-VP24
  • LT-PERMET-96
  • LT-PERMET-VP96
  • LudgerTag 2-AB Glycan Labelling Kit, sodium cyanoborohydride (24 samples)
  • LudgerTag 2-AB Glycan Labelling Kit, picoline borane (24 samples)
  • LudgerTag 2-AB Glycan Labelling Kit, picoline borane (96 samples)
  • LudgerTag Procainamide Glycan Labelling Kit, sodium cyanoborohydride (24 samples)
  • LudgerTag Procainamide Glycan Labelling Kit, sodium cyanoborohydride (96 samples)
  • LudgerTag Procainamide Glycan Labelling Kit, picoline borane (24 samples)
  • LudgerTag 2-AA Glycan Labelling Kit, sodium cyanoborohydride (24 samples)
  • LudgerTag 2-AA Glycan Labelling Kit, picoline borane (24 samples)
  • LudgerTag Permethylation Kit (96 samples)
  • LudgerTag Permethylation Kit, without methyl iodide (96 samples)

Alternative enzymes

  • E-BG07
  • E-BG02
  • β(1-4)-galactosidase
  • β(1-3,4,6)-galactosidase






References:

1. Kobata, A. Use of endo- and exoglycosidases for structural studies of glycoconjugates. Anal Biochem 100: 1- 14 (1979).

2. Prime, S. J. Dearnley, A.M. Venton, R.B. Parekh and C.J. Edge. Oligosaccharide sequencing based on exo- and endoglycosidase digestion and liquid chromatographic analysis of the products. J Chromatogr A 720: 263-274 (1996)

3. Dwek, R.A. , C.J. Edge, D.J. Harvey, M.R. Wormald and R.B. Parekh. Analysis of glycoprotein-associated oligosaccharides. Ann Rev Biochem 62: 65-100.