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.
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
Other Products
Fastin™ – Elastin assay kit
Product Info
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Product Info
Introducing the Fastin Assay Kit: Your Straightforward Solution for Elastin Quantification! Our user-friendly kit utilizes a dye-based method to measure elastin from in-vivo and in-vitro sources. It can be used to quantify various elastin forms, spanning from immature tropoelastin to mature, ‘insoluble’ elastin fibers.
Colorimetric Detection (513nm) (Endpoint)
Understanding Elastin: The Key to Tissue Flexibility
Tissues like lungs and arteries must maintain the ability to stretch and recoil repeatedly throughout an organism’s life. Elastin, a mature protein, is responsible for this elasticity and is usually present as insoluble fibers within the ECM. During development, these fibers are initially formed from a soluble precursor called tropoelastin.
What is the Fastin Assay?
The Biocolor Fastin assay is a user-friendly, dye-based means of quantifying elastins derived from both in-vivo and in-vitro sources. A variety of elastin forms can be assayed, from immature tropoelastin to mature ‘insoluble’ elastin fibres.
Further information on how the assay works can be found on the ‘Mode of Action‘ tab.
A list of suggested sample types can be found under the ‘Assay Specification‘ tab.
How does the Fastin assay detect Elastin?
The Fastin Dye Reagent contains an elastin-binding synthetic porphyrin, TPPS (5,10,15,20-tetraphenyl- 21H,23H-porphine). This affinity of TPPS for elastin was first observed when used as a ‘vital stain’ on live animals. Most tissues took up the dye initially but only elastin retained the TPPS molecules over time. [Winkelman, J. (1962), Cancer Res. 22, 589-596; Winkelman, J & Spicer, S. (1962), Stain Technol. 37, 303-305].
It has been proposed that the elastin binding of TPPS may be due to the retention of the acidic dye (which contains four charged sulfate groups) by the basic amino acid side chain residues of elastin.
How does the Fastin assay work?
Step 1. Incubation of samples containing soluble elastin with the Fastin Dye Reagent causes an elastin-dye complex to form. This insoluble complex then precipiates.
Step 2. Dye-labelled elastin is then isolated by centrifugation and the unbound dye removed. Elastin-bound dye is then eluted and measured spectrophotometrically.
Step 3. The elastin content of unknown samples can be calculated by comparison against a calibration curve prepared using a standard comprising water-soluble elastin (supplied with the kit).
Assay range
50 – 500µg/ml
Limit of Detection
50µg/ml
Detection Method
Colorimetric Detection (513nm) (Endpoint)
Measurements per kit
110 in total (allows a maximum of 48 samples to be run in duplicate alongside a standard curve).
Suitable Samples
In-vivo: tissues and fluids. Insoluble elastin will first require conversion to water soluble α-elastin using the oxalic acid reagents and extraction protocol supplied with the kit.
In-vitro: Elastin produced by cells during 2D/3D cell culture. NB elastin in conditioned cell media is typically below the detection limit of the kit.
Precautions
This kit is designed for research use only. Not for use in diagnostic procedures. Kit requires access to a centrifuge, heated water bath or block, as well as a spectrophotometer or colorimeter capable of absorbance detection at 513nm. Specific sample preparation protocols may require customer to provide further reagents, consult assay manual for further information.
Fastin elastin kit contents:
1. Dye Reagent (1x110ml)
2. α-elastin Reference Standard (1x5ml, 1.0 mg/ml soluble Bovine elastin)
6. 1.5ml micro-centrifuge tubes for dye-labelling reaction.
7. Assay kit manual
NB: Additional reagents may be required for sample preparation prior to assay. Consult manual or contact us for further details.
Document
Introducing the Fastin Assay Kit: Your Straightforward Solution for Elastin Quantification! Our user-friendly kit utilizes a dye-based method to measure elastin from in-vivo and in-vitro sources. It can be used to quantify various elastin forms, spanning from immature tropoelastin to mature, ‘insoluble’ elastin fibers. Colorimetric Detection (513nm) (Endpoint)
Purification and enrichment of intact urinary exosomes for functional studies
Isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias.
Versatile urine input volumes
No phenol extractions, Proteinase K treatment, nor carrier RNA required
No time-consuming ultracentrifugation, filtration nor special syringes are required
No precipitation reagents nor overnight incubation required
Compatible with urine from most species
Pure exosomes are purified and are free from any other RNA-binding proteins
Purification is based on spin column chromatography that uses Norgen’s proprietary resin
The purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
Norgen’s Exosome Purification and RNA Isolation Kits constitute all-in-one systems for the purification of exosomes and the subsequent isolation of exosomal RNA from different urine sample volumes. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. These kits are designed to isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias. These kits provide a clear advantage over other available kits in that they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, the kit allows the user to elute into a flexible elution volume ranging from 50 µL to 100 µL. The purified RNA is free from any protein-bound circulating RNA and of the highest integrity. The purified RNA can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.
Urine Exosome Purification and RNA Isolation Mini Kit
For sample volumes ranging from 250 µL to 1 mL.
Urine Exosome Purification and RNA Isolation Midi Kit
For sample volumes ranging from 2 mL to 10 mL.
Urine Exosome Purification and RNA Isolation Maxi Kit
All sizes, including miRNA and small RNA (< 200 nt)
Elution Volume
50-100 μL
Time to Complete 10 Purifications
35 – 40 minutes
Average Yields*
Variable depending on specimen
*Please check page 4 of the product insert for the average yields and the common RNA quantification methods.
Storage Conditions and Product Stability All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.
Important Note Urine samples stored at -70°C, -20°C or at 4°C will develop some precipitation due to the aggregation of some of the highly abundant proteins in urine. Eliminating these precipitates using centrifugation or filtration may cause the loss of exosomes. Furthermore, these precipitates may affect the quality of the purified nucleic acid. We recommend the use of Norgen’s Urine Preservative when collecting urine samples, which is designed for the preservation of nucleic acids and proteins in fresh urine samples at ambient temperatures. The components of the Urine Preservative allow samples to be stored for over 2 years at room temperature with no detected degradation of urine DNA, RNA or proteins. Norgen’s Urine Preservative is available as a liquid format in Norgen’s Urine Preservative Single Dose Ampules, as well as in a dried format in Norgen’s Urine Collection and Preservation Tubes.
AchE Inhibitor Screening Kit (Colorimetric) provides a rapid, simple, sensitive, and reliable test suitable for high-throughput screening of AchE inhibitors.
ACETYLCHOLINESTERASE (EC 3.1.1.7, AChE), also known as RBC cholinesterase, is found primarily in the blood and neural synapses. AChE catalyzes the hydrolysis of the neurotransmitter acetylcholine into choline and acetic acid, a reaction necessary to allow a cholinergic neuron to return to its resting state after activation. Inhibition of the enzyme leads to acetylcholine accumulation, hyperstimulation of nicotinic and muscarinic receptors, and disrupted neurotransmission. AChE inhibition is an important target for the management of Alzheimer’s disease. In addition to Alzheimer’s disease, AChE inhibitors have been useful in the diagnosis or treatment of diseases such as glaucoma, myasthenia gravis, bladder distention, and more.
The Attogene Acetylcholinesterase Inhibitor Assay is based on an improved Ellman method, in which thiocholine produced by the action of acetylcholinesterase forms a yellow color with 5,5′-dithiobis(2-nitrobenzoic acid). The intensity of the product color, measured at 412nm, is proportionate to the enzyme activity in the sample.
Document
The Acetylcholinesterase Assay Kit provides a convenient method for the detecting AChE activity and screening for inhibitors. The kit uses DTNB to quantify the thiolcholine produced from the hydrolysis of acetylthiolcholine by AChE.
