N-acetyl-ß-D-glucosaminidase
Spectrophotometric method

This kit procedure incorporates the VRA-GlcNAc substrate and provides an alternative to MNP-GlcNAc kit when a spectrophotometric measurement is required. The released colour is stable at 490nm and this avoids the use of a calibrant. The enzyme reaction is carried out at saturation and this together with the high extinction coefficient at 490nm (E 38,600, pH 9.5). of the chromophore at alkaline pH gives this high sensitivity. The procedure has been used mainly for the assay of urine from children where samples are often dilute and high sensitivity is required (see references).

Assay Procedure

Random urine sample (50µl) is added to substrate solution (700µl) and incubated for 30 minutes at 37°C. The reaction is stopped by the addition of buffer pH 10. the colour released is measured at 490nm.

The activity of the enzyme in µmoles/min/L of chromophore released is then calculated according to the equation:

available in 50test and 100test kits

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