Exosomal RNA Isolation Kit (for the extraction of RNA from EVs that have been isolated using IZON’s qEV columns)
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Detail
Overview
Extract high quality & quantity total RNA including miRNA
No phenol step required; isolate all RNA in one fraction
Bind & elute all RNA irrespective of size or GC content, without bias
Very sensitive & linear down to a few cells without the need for carrier RNA
Isolate from a wide variety of specimens
Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
Available in a variety of formats to suit your needs
Purification is based on spin column chromatography that uses Norgen’s resin separation matrix
Norgen’s Exosomal RNA Isolation Kit (for the extraction of RNA from EV’s that have been isolated using IZON’s qEV columns) constitutes an all-in-one system for the isolation of exosomal RNA from exosomes previously isolated using IZON’s qEV columns or concentrated using IZON’s EV Concentration kit. This kit allows for the isolation of RNA from intact extracellular vesicles (EVs) where the purification is based on spin column chromatography that employs Norgen’s proprietary resin.
The kit is designed to isolate all sizes of extracellular vesicle RNA, including microRNA. The kit provides a clear advantage over other available kits in that it does 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 of the highest integrity, and can be used in a number of downstream applications including real time PR, reverse transcription PCR, Nothern blotting, RNase protection and primer extension, and expression array assays.
Details
Storage Conditions and Product Stability All buffers should be kept tightly sealed and stored at room temperature (15–25°C). 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.
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.
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.
Collagen is a fundamental component of the extracellular matrix, and the predominant protein in animals, constituting around 30% of total protein mass. A glycoprotein, it is well known for its triple helical structure. This is formed from three polypeptide α-chains with Gly-X-Y repeating residues (Gly for Glycine, X for proline, and Y for hydroxyproline).
Types of Collagen
Over 28 types of collagens have been identified, with Type I collagen being the most abundant. It’s prevalent in ligaments, tendons, skin, and bone tissue. Its mature, insoluble form grants it remarkable strength, making it vital for the mobility of organisms. Collagen also has biochemical functions, influencing cell growth, proliferation, and differentiation.
This version of the kit is designed to detect and measure INSOLUBLE forms of collagen. Chose our Sircol 2.0 collagen kit if you need to analyse SOLUBLE collagen.
Applications of Collagen
Collagen, with its diverse properties, finds utility in various industries. It plays a role in medicine for wound healing and has an expanding role in tissue engineering and cell culture for biomedical purposes. It’s gaining popularity in the cosmetic industry for skin rejuvenation and is used in chemical formulations and the food industry as a functional food supplement and additive.
How does the Sircol assay detect collagen?
Sircol dye reagent contains Sirius Red – a linear anionic dye with sulphonic acid side chain groups. Under assay conditions the Sircol dye binds the basic groups of soluble collagen molecules. Maximal binding occurs in collagens possessing intact triple helix organisation as the highly ordered Gly-X-Yn helical structure of tropocollagen further contributes to dye binding. This results in a high degree of dye-collagen specificity. Affinity is progressively reduced during heat denaturation 4ºC due to the unwinding of the triple helix and formation of random chains.
Overview of the Sircol assay process:
Step 1. Samples being assayed for insoluble collagen must first undergo a 2-3 hour pre-treatment with Sircol Fragmentation reagent. This converts insoluble collagen into water-soluble gelatin can then be assayed.
Step 2. Addition of Sircol Dye Reagent to these pre-treated insoluble collagen samples results in the formation of a denatured collagen-dye complex. This complex then precipitates during the dye incubation period and is subsequently isolated by centrifugation, followed by washing to remove unbound dye. The Denatured collagen-bound dye is then eluted and measured spectrophotometrically.
Step 3. The insoluble collagen content of unknown samples is quantified by comparison against a calibration curve prepared using a the denatured collagen standard supplied with the kit.
Assay range
100 – 1000 µg/ml
Limit of Detection
100µg/ml
Detection Method
Colorimetric Detection (556nm) (Endpoint)
Measurements per kit
110 in total (allows a maximum of 46 samples to be run in duplicate alongside a standard curve).
Suitable Samples
The assay can be used to assess the rate of production of newly laid down collagen fibres during periods of rapid growth, development, tissue repair, remodeling and wound healing. Sources of material includes tissues, bone and calcified tissue.
*Insoluble collagens must be converted into soluble form prior to assay. Instructions and regents are provided with the kit., depending on sample this will require prior salt/acid/acid-pepsin extraction.
**non-mammalian collagens may result in a reduced limit of detection. We recommend use of an assay standard matched to the species under assay.
Many customers have found that the straightforward sample processing and analysis of Sircol make it a good alternative to conventional hydroxyproline analysis.
Precautions
This kit is designed for research use only. Not for use in diagnostic procedures. Kit requires access to a centrifuge, water bath / heated block, as well as a spectrophotometer/colorimeter capable of absorbance detection at 556nm. Specific sample preparation protocols may require customer to provide further reagents, consult assay manual for further information.
Sircol Insoluble Collagen kit contents:
1. Sircol Dye Reagent (1x110ml)
2. Denatured Collagen Reference Standard (1x5ml, 1.0mg/ml)
3. Acid-Salt Wash Reagent (1x20ml)
4. Fragmentation Reagent (1x110ml)
5. Alkali Reagent (1x110ml)
6. 2ml screw-cap tubes for preparation of samples.
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
As collagens mature, they become increasingly crosslinked and insoluble – characteristics necessary for key biophysical role that collagen plays in living organisms. Biocolor’s Sircol™ INSOLUBLE Collagen Kit is a dye-binding assay designed for accurate quantification and measurement such collagens. It is ideal for analyzing crosslinked / insoluble collagens from sources such as tissues, bone, and calcified tissue.
