New England Biolabs
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  • Glycosidases Double Digest Chart

    NEB's glycosidases are provided with 10X buffer to ensure optimal activity. Using more than one glycosidase simultaneously is a common timesaving procedure. Selecting the best buffer to provide reaction conditions that optimize enzyme activity is an important consideration. The following chart will help in selecting the optimal conditions for double digests.

    NEB # P0734 P0732 P0721 P0702
    P0703
    P0733 P0724 P0731 P0726 P0730 P0729 P0727 P0720 P0728 P0704
    P0705
    P0704
    P0705
    G7 G7 G7 S G7 G7 G7 G7 G7 G7 G7 G7  G7 G7
    P0702
    P0703
    S S S G5 G5 S S S S S S G5 S S
    P0733 G7 G7 G7 G5 G7 G7 G7 G7 G7 G7 G7 G7 G7 G7
    P0726 G6 G6 G6 S G7 G6 G6 G2 G6 G6 G6 G6 G6 G7
    P0730 G6 G6 G6 S G7 G6 G6 G6 G4 G6 G6 G6 G6 G7
    P0731 G6 G6 G6 S G7 G6 G6 G6 G6 G6 G6 G6 G6 G7
    P0732 G6 G1 G6 S G7 G6 G6 G6 G6 G6 G6 G6 G6 G7
    P0734 G7 G6 G6 S G7 G6 G6 G6 G6 G6 G6 G6 G6 G7
    P0721 G6 G6 G2 S G7 G6 G6 G6 G2 G6 G6 G6 G6 G7
    P0729 G6 G6 G6 S G7 G6 G6 G6 G6 G6 G6 G6 G6 G7
    P0727 G6 G6 G6 S G7 G6 G6 G6 G6 G6 G2 G6 G6 G7
    P0720 G6 G6 G6 G5 G7 G6 G6 G6 G6 G6 G6 G1 G6 G7
    P0728 G6 G6 G6 S G7 G6 G6 G6 G6 G6 G6 G6 G4 G7
    P0724 G6 G6 G6 S G7 G4 G6 G6 G6 G6 G6 G6 G6 G7

    Notes

    P0702 = Endo H
    P0703 = Endo Hf
    P0704 = PNGaseF
    P0705 = PNGaseF (Glycerol Free)
    P0720 = Neuraminidase
    P0721 = β-N-Acetylhexosaminidasef
    P0724 = α1-2 Fucosidase
    P0726 = β1-3 Galactosidase
    P0727 = α1-6 Mannosidase
    P0728 = α2-3 Neuraminidase
    P0729 = α1-2,3 Mannosidase
    P0730 = β(1-4) Galactosidase
    P0731 = α(1-3,6) Galactosidase
    P0732 = β-N-Acetylglucosaminidase
    P0733 = O-Glycosidase
    P0734 = α-N-Acetylgalactosaminidase

    S = Sequential digest under denaturing conditions
    (Heat kill and sequential digest with appropriate buffer for each enzyme)
    If BSA is required for one enzyme, include it in the double digest.

    Reaction Buffer Compositions

    1X G1 Reaction Buffer:
    50 mM Sodium Citrate (pH 6.0 @ 25°C).

    1X G2 Reaction Buffer:
    50 mM Sodium Citrate (pH 4.5 @ 25°C).

    1X G4 Reaction Buffer:
    50 mM Sodium Citrate (pH 6.0 @ 25°C)
    and 100 mM NaCl.
    1X G5 Reaction Buffer:
    50 mM Sodium Citrate (pH 5.5 @ 25°C).

    1X G6 Reaction Buffer:
    50 mM Sodium Acetate (pH 5.5 @ 25°C)
    and 5 mM CaCl2.

    1X G7 Reaction Buffer:
    50 mM Sodium Phosphate (pH 7.5 @ 25°C).