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Cellafluor Reagent

Product code: R-CELLFLR
€161.00

100 assays (manual) / 1000 assays (auto-analyser) / 1000 assays (microplate)

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Available for shipping

Content: 100 assays (manual) / 1000 assays (microplate) / 1000 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, Microplate, Auto-analyser
Detection Method: Fluorescence
Wavelength (nm): 365 (ex), 450 (em)
Reproducibility (%): ~ 3%

Cellafluor is a fluorimetric reagent that can be used for the assay of cellulase (endo-1,4-β-glucanase). The reagent consists of 2 components:

1)      4,6-O-Benzylidene-4-methylumbelliferyl-β-cellotrioside

2)      β-glucosidase (Thermotoga maritima)

Upon incubation with a cellulase (endo-1,4-β-glucanase) enzyme, the functionalised trisaccharide is cleaved and the ancillary β-glucosidase can act to hydrolyse the fragment containing the 4-methylumbelliferyl group. The rate of formation of 4-methylumbelliferyl is directly proportional to the cellulase (endo-1,4-β-glucanase) activity. The assay reaction is terminated and full fluorescence obtained from the fluorophore by the addition of Tris buffer.

Display all of our reagent mixture products.

Documents
Certificate of Analysis
Safety Data Sheet
Booklet
Publications
Megazyme publication
Quantitative fluorometric assay for the measurement of endo-1,4-β-glucanase.

Mangan, D., McCleary, B. V., Liadova, A., Ivory, R. & McCormack, N. (2014). Carbohydrate Research, 395, 47-51.

There is a growing demand for research tools to aid the scientific community in the search for improved cellulase enzymes for the biofuel industry. In this work, we describe a novel fluorometric assay for cellulase (endo-1,4-β-glucanase) which is based on the use of 4,6-O-benzylidene-4-methylumbelliferyl-β-cellotrioside (BzMUG3) in the presence of an ancillary β-glucosidase. This assay can be used quantitatively over a reasonable linear range, or qualitatively as a solution screening tool which may find extensive use in the area of metagenomics.

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Megazyme publication
Novel substrates for the measurement of endo-1,4-β-glucanase (endo-cellulase).

McCleary, B. V., Mangan, D., Daly, R., Fort, S., Ivory, R. & McCormack, N. (2014). Carbohydrate Research, 385, 9-17.

A specific and sensitive substrate for the assay of endo-1,4-β-glucanase (cellulase) has been prepared. The substrate mixture comprises benzylidene end-blocked 2-chloro-4-nitrophenyl-β-cellotrioside (BzCNPG3) in the presence of thermostable β-glucosidase. Hydrolysis by exo-acting enzymes such as β-glucosidase and exo-β-glucanase is prevented by the presence of the benzylidene group on the non-reducing end D-glucosyl residue. On hydrolysis by cellulase, the 2-chloro-4-nitrophenyl-β-glycoside is immediately hydrolysed to 2-chloro-4-nitrophenol and free D-glucose by the β-glucosidase in the substrate mixture. The reaction is terminated and colour developed by the addition of a weak alkaline solution. The assay procedure is simple to use, specific, accurate, robust and readily adapted to automation. This procedure should find widespread applications in biomass enzymology and in the specific assay of endo-1,4-β-glucanase in general.

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Safety Information
Symbol : Not Applicable
Signal Word : Not Applicable
Hazard Statements : Not Applicable
Precautionary Statements : Not Applicable
Safety Data Sheet
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