The Glucuronoyl Esterase Assay Kit provides a simple robust method for the measurement of glucuronoyl esterase. The assay kit employs a novel colourimetric reagent, termed GEUX3 that is highly specific for glucuronoyl esterase.
Detail
K-GEUX3
SKU: 700004294
100 assays per kit
Content:
100 assays per kit
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:
Glucuronoyl Esterase
Assay Format:
Spectrophotometer, Auto-analyser
Detection Method:
Absorbance
Wavelength (nm):
400
Signal Response:
Increase
Reproducibility (%):
~ 3%
Total Assay Time:
10 min
Application examples:
Fermentation broths, industrial enzyme preparations and biofuels research.
Method recognition:
Novel method
The Glucuronoyl Esterase Assay Kit provides a simple robust method for the measurement of glucuronoyl esterase. The assay kit employs a novel colourimetric reagent, termed GEUX3 that is highly specific for glucuronoyl esterase.
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)
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 1 x 1010 AAV particles as determined by qPCR
AAV Vector Serotype
Any
Average Recovery
> 80%
Input Type
Cells, media, or mixed
Input Volume
8 mL – 45 mL
Minimum Elution Volume
1 mL
Time to Complete 4 Purifications
2 – 2.5 hours
Storage Conditions and Product Stability DNAse I and RNAse A 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 1 year 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
