Cellulase Assay Kit (CellG5 Method)

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The CellG5 assay reagent for the measurement of endo-cellulase (endo-1,4-β-glucanase) contains two components; 1) 4,6-O-(3-Ketobutylidene)-4-nitrophenyl-β-D-cellopentaoside (BPNPG5) and 2) thermostable β-glucosidase. The ketone blocking group prevents any hydrolytic action by the β-glucosidase on BPNPG5. Incubation with an endo-cellulase generates a non-blocked colourimetric oligosaccharide that is rapidly hydrolysed by the ancillary β-glucosidase. The rate of formation of 4-nitrophenol is therefore directly related to the hydrolysis of BPNPG5 by the endo-cellulase. The reaction is terminated and the phenolate colour is developed on addition of Tris buffer solution (pH 9.0).

Detail

K-CellG5-4V

SKU: 700004272

Content:(K-CellG5-4V)
120 / 240 assays (manual) / 480 assays (auto-analyser) or 
(K-CellG5-2V)
60 / 120 assays (manual) / 240 assays (auto-analyser)
Shipping Temperature:Ambient
Storage Temperature:Short term stability: 2-8oC,
Long term stability: See individual component labels
Stability:> 2 years under recommended storage conditions
Analyte:endo-Cellulase
Assay Format:Spectrophotometer, Auto-analyser
Detection Method:Absorbance
Wavelength (nm):400
Signal Response:Increase
Limit of Detection:1.2 x 10-3 U/mL
Reproducibility (%):~ 3%
Total Assay Time:10 min
Application examples:Fermentation broths, industrial enzyme preparations and biofuels research.
Method recognition:Novel method

The K-CellG5-2V pack size has been discontinued (read more).

Cellulase Activity Assay Kit.

The CellG5 assay reagent for the measurement of endo-cellulase (endo-1,4-β-glucanase) contains two components; 1) 4,6-O-(3-Ketobutylidene)-4-nitrophenyl-β-D-cellopentaoside (BPNPG5) and 2) thermostable β-glucosidase. The ketone blocking group prevents any hydrolytic action by the β-glucosidase on BPNPG5.  Incubation with an endo-cellulase generates a non-blocked colourimetric oligosaccharide that is rapidly hydrolysed by the ancillary β-glucosidase. The rate of formation of 4-nitrophenol is therefore directly related to the hydrolysis of BPNPG5 by the endo-cellulase. The reaction is terminated and the phenolate colour is developed on addition of Tris buffer solution (pH 9.0).

The CellG5 assay represents a huge step forward in the methodology for the measurement of cellulase that traditionally relied on substrates such as CM-cellulose, Avicel, cellooligosaccharides, filter paper or dyed polysaccharides including CMC Congo red or cellulose azure.

View Cellulase Activity Assay Protocol.

View our complete list of assay kits for enzyme activities.

Scheme-K-CellG5-2V CellG5 Megazyme

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