The rate of hydrolysis increases with buffer pH and contributes to less-efficient crosslinking in less-concentrated protein solutions. The half-life of hydrolysis for NHS-ester compounds is 4 to 5 h at pH 7.0 and 0°C. This half-life decreases to 10 mins at pH 8.6 and 4°C. The extent of NHS-ester hydrolysis in aqueous solutions free of primary amines can be measured at 260 to 280 nm, because the NHS byproduct absorbs in that range.
What is hydrolysis rates for NHS Ester functionalized dyes?
NHS ester tends to hydrolyze over the time in water.
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