029150 Phenylanine Deaminase Reagent

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.

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).

Checkit Go enables you to easily measure the volumes dispensed by a robot or multi-channel pipette, in under 10 seconds. These models measure up to 8 channels at a time and are offered in volume ranges covering 2μL to 50μL.

Please choose the model that matches your volume range:

Model	Volume Range
5 μL Volumetric Verification Kit	2-5 μL
10 μL Volumetric Verification Kit	4-10 μL
20 μL Volumetric Verification Kit	8-20 μL
50 μL Volumetric Verification Kit	20-50 μL

Checkit Go enables you to easily measure the volumes dispensed by a robot or multi-channel pipette, in under 10 seconds. These models measure up to 8 channels at a time and are offered in volume ranges covering 2μL to 50μL.

Please choose the model that matches your volume range:

Model Volume Range
5 μL Volumetric Verification Kit 2-5 μL
10 μL Volumetric Verification Kit 4-10 μL
20 μL Volumetric Verification Kit 8-20 μL
50 μL Volumetric Verification Kit 20-50 μL

Overview

• Columns bind proteins of interest while endotoxins flow through
• Reduce endotoxin levels to less than 0.01 EU/µg of protein
• Proteins are desalted
• Greater than 95% protein recovery
• Concentrate protein samples and remove detergents at the same time
• Effectively remove a wide range of detergents including SDS, Triton® X-100, CHAPS, NP-40, and Tween 20
• Purification is based on spin column chromatography that uses Norgen’s resin separation matrix

The ProteoSpin™ Total Protein Concentration, Detergent Clean-Up and Endotoxin Removal Kits are designed for the rapid removal of endotoxins from previously purified proteins or peptides, with protein recoveries of > 95% being achieved. Endotoxins, also known as lipopolysaccharides, are cell-membrane components of Gram-negative bacteria such as E. coli. Endotoxins liberated by Gram-negative bacteria are frequent contaminations of protein solutions derived from bioprocesses. Due to the high toxicity of endotoxins in vivo and in vitro, their removal from protein preparations is often necessary prior to the use of the protein in downstream applications. These kits efficiently reduce endotoxin levels to ≤ 0.01 EU/mg of protein, using spin column chromatography based on Norgen’s proprietary resin as the separation matrix. These kits are highly efficient in removing many different salts commonly used in the laboratory including, but not limited to, MgCl₂, NaCl, KCl, CaCl₂, LiCl and CsCl. The purified protein samples can be used in a number of downstream applications including sequencing, cloning, and in vitro and in vivo introduction into cells and organisms for various purposes. The simultaneous removal of salts while concentrating a dilute protein solution makes the kit a convenient method for preparing proteins before running many downstream applications such as SDS-PAGE, isoelectric focusing, X-ray crystallography, NMR spectroscopy, mass spectroscopy and other applications.

Mini Kit

The ProteoSpin™ Total Protein Concentration, Detergent Clean-Up and Endotoxin Removal Mini Kit is designed for the rapid removal of endotoxins from up to 200 μg of previously purified proteins or peptides, with protein recoveries of > 95% being achieved.

Maxi Kit

The ProteoSpin™ Total Protein Concentration, Detergent Clean-Up and Endotoxin Removal Maxi Kit is designed for the rapid removal of endotoxins from up to 4 mg of previously purified proteins or peptides, with protein recoveries of > 95% being achieved.

Details

Supporting Data

Figure 1 / 15

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Kit Specifications
Maximum Protein Input	4 mg
Minimum Protein Input	0.25 mg
Maximum Column Volume Input	18 mL
Molecular Weight of Recovered Proteins	No Molecular Weight cut-off
Final Endotoxin Levels	≤ 0.01 EU/μg protein
Protein Recovery	90-95%
% Detergent Removal	90-95%
Detergents that can be Removed	Including Triton® X-100, CHAPS, NP-40 and Tween 20
Elution Volume	2-4 mL
Time to Complete 10 Purifications	30-40 minutes

Storage Conditions
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.

Component	Cat. 22800 (25 preps)	Cat. 22200 (4 preps)
Binding Buffer J	8 mL	8 mL
Binding Buffer N	4 mL	20 mL
Wash Solution M	50 mL	130 mL
Wash Solution CIP	20 mL	60 mL
Wash Solution N	30 mL	130 mL
Wash Solution NIP	20 mL	60 mL
Elution Buffer G	6 mL	20 mL
Endotoxin Removal Solution	1.5 mL	1.5 mL
Protein Neutralizer EF	2 mL	2 mL
Maxi Spin Columns (assembled with Collection Tubes)	–	4
Mini Spin Columns	25	–
Collection Tubes	25	–
Maxi Spin Columns (assembled with Collection Tubes)	4
Elution Tubes (1.7 mL)	25	–
Elution Tubes (50 mL)	4	–
Product Insert	1	1