PEG13-bis(Amino-Tri-(Propargyl-PEG8-ethoxymethyl)-methane)

Overview

• Rapid AAV DNA extraction and quantification can be completed in 2 hours 
• Universal qPCR assay targeting the AAV2 ITR enables quantification of most AAV vector constructs
• Purified DNA standard enables easy comparison of data between different lab groups
• Non-plasmid based standard prevents quantification artifacts due to insufficient melting of plasmid sequences
• DNAse digestion step aids in eliminating non-viral DNA
• Minimal sample handling for maximum AAV DNA recovery

Norgen’s AAV Quantification Kit is designed for the detection of adeno-associated virus (AAV) inverted terminal repeat (ITR) sequences in a real-time PCR based on the use of TaqMan® technology. A purified standard based solely on the AAV2 ITR sequence purified using Norgen’s proprietary silicon carbide technology simplifies the generation of a reliable standard curve for AAV quantification. Avoidance of a plasmid based standard eliminates problems associated with efficient melting of the ITR sequence due to coupling of the ITR to the much longer plasmid sequence, as well as variability due to rearrangements/duplications/deletions of the recombination prone ITR. An easy and rapid method for viral DNA extraction simplifies the step of obtaining AAV DNA while simultaneously eliminating contaminating non-encapsidated DNA.  Norgen’s AAV Quantification Kit can facilitate pre-clinical studies that require accurate vector titration as well as interlab comparisons of vector quantities. This kit is designed for research use only and not for use in diagnostic procedures.

Details

Supporting Data

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Storage Conditions and Product Stability
All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots. All reagents can be stored for 1 year at -20°C without showing any reduction in performance.

Component	Cat. 63800 (24 rxns)
Enzyme Incubation Buffer A	500 µL
DNase I	25 µL
DNase Stop Solution	100 µL
2X PCR Mastermix	350 µL
AAV Primer & Probe Mix	90 µL
AAV Standard	6 µL
Nuclease-Free Water (Negative Control)	1.25 mL
Product Insert	1

K-MBG4

SKU: 700004319

100 assays (manual) / 400 assays (auto-analyser)

Content:	(Malt β-glucanase) 100 assays (manual) / 400 (auto-analyser) Or (Lichenase) 100 / 200 assays (manual) / 330 (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:	β-Glucanase/Lichenase
Assay Format:	Spectrophotometer, Auto-analyser
Detection Method:	Absorbance
Wavelength (nm):	400
Signal Response:	Increase
Limit of Detection:	(Malt β-glucanase) 4.3 x 10-4 U/mL Or (Lichenase) 9.1 x 10-5 U/mL
Reproducibility (%):	~ 3%
Total Assay Time:	(Malt β-glucanase) ~ 20 min Or (Lichenase) ~ 10 min
Application examples:	Crude malt extracts, industrial enzyme preparations.
Method recognition:	Novel method

The MBG4 reagent contains a single substrate, namely 4,6-O-benzylidene-2-chloro-4-nitrophenyl-β-(3¹-β-D-cellotriosyl-glucoside) (BCNPBG4). The benzylidene acetal group prevents any hydrolytic action by exo-acting hydrolytic enzymes such as β-glucosidase or cellobiohydrolase.

Mixed linkage β-glucanase (endo-1,3:1,4-β-glucanase) / lichenase (EC 3.2.1.73) acts specifically to release 2-chloro-4-nitrophenol (CNP) from this substrate. The rate of release of CNP is directly related to the β-glucanase/lichenase activity in a sample. The reaction is terminated and the phenolate colour is developed on addition of Tris buffer solution (pH = 10.0).

Note that the substrate is not hydrolysed by β-glucosidase or cellobiohydrolase. The substrate can be hydrolysed by certain endo-cellulases (e.g. Trichoderma sp.) but this does not result in an increase in absorbance.

Discover more assay kits for enzyme activity measurement.

Data calculators are located in the Documents tab.

Advantages

• Very cost effective 
• All reagents stable for > 2 years 
• Specific for endo-1,3:1,4-β-glucanase/lichenase 
• Simple, convenient, rapid assay 
• Well suited to automation 
• Malt flour standard and lichenase standard included

The MBG4 reagent contains a single substrate, namely 4,6-O-benzylidene-2-chloro-4-nitrophenyl-β-(31-β-D-cellotriosyl-glucoside) (BCNPBG4). The benzylidene acetal group prevents any hydrolytic action by exo-acting hydrolytic enzymes such as β-glucosidase or cellobiohydrolase.

Description

Formaldehyde is an organic compound with the formula CH2O. It is mainly used in the production of industrial resins but has been found to be used as a preservative, disinfectant and biocide. Because of its toxicity and volatility formaldehyde poses a significant risk to human health. Attogene test uses the property of formaldehyde to react with the Formaldehyde Reaction Powder (FRP) to form a purple-red tetrazine. The formaldehyde concentration is measured by visual comparison of the reaction with the color scale derived from the Color Card.

Measuring range / color- Number of scale graduation	Number of determinations
0.1 – 0.25 – 0.4 – 0.6 – 0.8 – 1.0 – 1.5 mg/l HCHO	100

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Where formaldehyde is found

Formaldehyde is found in:

• Resins used in the manufacture of composite wood products (i.e., hardwood plywood, particleboard and medium-density fiberboard)
• Building materials and insulation
• Household products such as glues, permanent press fabrics, paints and coatings, lacquers and finishes, and paper products
• Preservatives used in some medicines, cosmetics and other consumer products such as dishwashing liquids and fabric softeners
• Fertilizers and pesticides

It is a byproduct of combustion and certain other natural processes, and so is also found in:

• Emissions from un-vented, fuel burning appliances, like gas stoves or kerosene space heaters.
• Cigarette smoke.

White Paper from Nix Color Sensor using Attogene’s Formaldehyde Kit: Nix Color Sensor & Attogene Formaldehyde Kit

Attogene test uses the property of formaldehyde to react with the Formaldehyde Reaction Powder (FRP) to form a purple-red tetrazine. The formaldehyde concentration is measured by visual comparison of the reaction with the color scale derived from the Color Card.