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).
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.
AAV Purification from any input – cell fraction or media fraction
High AAV recovery, up to 90%
No specialized equipment needed
Purification from a variety of AAV serotypes (including AAV6 and AAV9)
Yields highly active AAV for in vivo and in vitro experiments
Purification is based on spin column chromatography that uses Norgen’s resin separation matrix
Recombinant adeno-associated virus (AAV) vectors are highly promising tools for both in vitro and in vivo gene transfer. Norgen’s AAV Purification Kits provide fast and simple procedures for concentrating and purifying AAV vectors from cell lysate and cell culture media. Purification is based on precipitation onto Norgen Biotek’s proprietary resin. Contaminating cellular debris is largely removed from the sample via a centrifugation step, while contaminating DNA and RNA is reduced using enzymatic digestion. AAV vector purified in this manner is highly active for use in in vitro and in vivo transduction experiments.
AAV Purification Kit
Norgen’s AAV Purification Kit contains sufficient materials for 15 preparations (33.5 mL per prep of supernatant (SN) or a total of 500 mL of supernatant input). Approximately 1 mL of cell pellet can be purified per prep, up to a maximum of 15 mL of cell pellet in total for the entire kit. Up to 33X sample concentration.
AAV Purification Mini Kit
Each spin column is able to concentrate and purify AAV from 0.5-8 mL of cell pellet, cell culture media, or cells and culture media mixed together. Up to 50X sample concentration. AAV vector purified in this manner is highly active for use in in vitro transduction experiments, and is eluted into a small volume (200 µL). Preparation time for 4 samples is 1.5 hours, with 45 minutes of hands-on time.
AAV Purification Midi Kit
Each spin column is able to concentrate and purify AAV from 8 mL up to 45 mL of input consisting of cell pellet, cell culture media, or cells and culture media mixed together. Up to 50X sample concentration. AAV vector purified in this manner is highly active for use in in vitro transduction experiments, and is eluted into a small volume (1 mL). The kit may be used to purify up to 8 x 25 mL or 4 x 45 mL of samples using the included columns. Preparation time for 4 samples is approximately 2 to 2.5 hours, with 1.5 hours of hands on time.
AAV Purification Maxi Kit (Slurry Format)
Each spin column is able to concentrate and purify AAV from 45 mL to 90 mL of input consisting of cell pellet, cell culture media, or cells and culture media mixed together. Up to 200X sample concentration. AAV vector purified in this manner is highly active for use in in vitro transduction experiments, and is eluted into a small volume (1-10 mL) using the optional concentration step. The kit may be used to purify up to 1 x 900 mL samples or 10 x 45-90 mL samples using the included columns. Preparation time for 1 x 900 mL sample is approximately 2.5 to 3.5 hours, with an optional concentration step requiring an additional 30 min.
At least 5 x 1010 AAV particles as determined by qPCR
AAV Vector Serotype
AAV6, AAV9 and others
Input Type
Cells, media
Input Volume (AAV supernatant)
1 – 33.5 mL SN per prep (500 mL SN in total)
Input Volume (AAV cell pellet)
1 mL cell pellet per prep (15 mL in total)
Time to Complete Purifications
2.5 to 4.5 hours with 1 hour hands on time
In vivo transduction
Yes
Storage Conditions and Product Stability
HL-SAN Nuclease should be stored at -20°C upon arrival. Elution Buffer O should be stored tightly capped at 4°C upon arrival. All other solutions should be kept tightly sealed and stored at room temperature. Once opened, the solutions should be stored at 4°C. This kit is stable for 2 years after the date of shipment.
Component
Cat. 66100 (15 preps)
Cat. 63200 (20 preps)
Cat. 63300 (4-8 preps)
Cat. 63250 (1-10 preps)
Lysis Buffer S
5.5 mL
5.5 mL
5.5 mL
20 mL
DNAse I
–
2 x 25 uL
2 x 25 uL
210 μL
RNAse A
–
60 μL
60 μL
240 μL
HL-SAN Nuclease
102 μL
–
–
–
Binding Buffer A
20 mL
4 mL
4 mL
2 x 8 mL
Purification Solution C
60 mL
–
–
–
Purification Solution D
130 mL
–
–
–
Wash Solution C
2 x 130 mL
60 mL
60 mL
3 x 60 mL
Slurry E
12.5 mL
–
–
2 x 14.5 mL
Elution Buffer O
66 mL
8.5 mL
8.5 mL
66 mL
Protein Neutralizer
4 mL
4 mL
4 mL
4 mL
Spin Columns
–
20
–
–
Mini Spin Columns
–
20
–
–
Midi Spin Columns (grey contents) with Collection Tubes
–
–
8
10
Midi Spin Columns (white contents) with Collection Tubes
–
–
8
–
Maxi Spin Columns (grey contents) with Collection Tubes
–
–
–
10
Maxi Spin Columns (white contents) with Collection Tubes
Simultaneous concentration, desalting and buffer exchange of total urinary proteins
No molecular weight cutoff allows for isolation of all sizes of proteins and peptides
Purification is based on spin column chromatography that uses Norgen’s resin separation matrix
These kits concentrate urine proteins while simultaneously removing salts, urea, and other urine contaminants. There is no molecular weight cut-off and therefore the columns capture total urinary proteins and peptides of all sizes making them ideal for biomarker discovery work, differential expression of proteins in various diseases, or other diagnostic research. The columns are convenient, rapid and easy to use and thus offer significant time savings over classic dialysis protocols. The resulting high-quality protein sample is concentrated and free from the original sample salts, thus preparing the sample conveniently for downstream proteomic applications including SDS-PAGE, 2D Gels, MALDI-TOF, LC/MS, LC/MS/MS, whole protein mass spectrometry, western blotting, protein microarrays, and more.
ProteoSpin™ Urine Protein Concentration Micro Kit
Each spin column is able to concentrate and desalt up to 200 μg of urine proteins. Twelve samples can be processed in 20 minutes.
ProteoSpin™ Urine Protein Concentration Midi Kit
The ProteoSpin™ Urine Protein Concentration Midi Kit provides a fast and simple procedure for concentrating dilute solutions of urine proteins from 1 to 5 mL inputs of urine. Each mini spin column is able to concentrate and desalt up to 3 mg of urine proteins in 30 minutes.
ProteoSpin™ Urine Protein Concentration Maxi Kit
The ProteoSpin™ Urine Protein Concentration Maxi Kit provides a fast and simple procedure for concentrating dilute solutions of urine proteins from 2 to 20 mL inputs of urine. Each maxi spin column is able to concentrate and desalt up to 4 mg of urine proteins in 45 minutes.
Storage Conditions All solutions should be kept tightly sealed and stored at room temperature. Once opened, the solution should be stored at 4°C. This kit is stable for 2 years after the date of shipment.
Component
Cat. 17400 (25 preps)
Cat. 52300 (10 preps)
Cat. 21600 (4 preps)
Wash Solution C
60 mL
60 mL
130 mL
Binding Buffer A
4 mL
4 mL
8 mL
Elution Buffer C
8 mL
30 mL
30 mL
Protein Neutralizer
4 mL
4 mL
4 mL
Micro Spin Columns
25
–
–
Midi Spin Columns (assembled with collection tubes)
–
10
–
Maxi Spin Columns (assembled with collection tubes)
