1 | Preparation of the reaction buffer, containing 6.1 mM CMP-NANA, 200 mM MES, pH 6.5. •Dissolve 40 mg CMP-NANA and 390.4 mg MES in 6 ml of water. •Adjust pH to 6.5. •Adjust to a final volume of 10 ml with water. | ||
2 | Setting up the sialylation reaction, containing 500 μg CMP-NANA, 100 mg ST3Gal-6, 1 mg IgG. Pipet the following solutions into a reaction tube: •125 μl reaction buffer •100 μl IgG (10 mg/ml, pH 6) •18 μl α-2,3-Sialyltransferase •7 μl water | ||
3 | Incubate the reaction at +37°C. | ||
4 | Stop the reaction by freezing the tube at -15 to -25°C. | ||
5 |
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Vial / Bottle | Label | Function / Description | Content |
---|---|---|---|
1 | α-2,3-Sialyltransferase rec. | Enzyme Storage buffer contains 50 mM MES, 200 mM NaCl, pH 6.4 at +4°C | Solution |