Excellent separation and purification of cytoplasmic and nuclear RNA
Convenient and fast spin column format
High quality and yield of RNA
Isolate full diversity of RNA (including microRNA) without phenol
Purified RNA is ready for any application including RT-PCR, qRT-PCR, RNA-Seq, arrays and more
Cytoplasmic RNA is free of DNA and ready for direct use in RT-PCR/qRT-PCR
Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix
This kit provides a rapid method for the isolation and purification of both cytoplasmic and nuclear RNA from cultured animal cells and small tissue samples. The kit can be used to isolate all sizes of RNA from the cytoplasmic and nuclear RNA fractions, including all small RNA species without any requirement for phenol. Included in the kit are sufficient reagents to perform either 50 cytoplasmic RNA preparations or 25 cytoplasmic and 25 nuclear RNA preparations. Ten samples can be processed in approximately 45 minutes. This kit is also available in a 100 prep size.
Background
In certain circumstances it is desirable to be able to isolate fractionated RNA as opposed to total RNA. For example, it may be preferable to isolate only mature, cytoplasmic RNA for some studies on expression profiling. Alternatively it may be desirable to isolate nuclear RNA in order to investigate and study pre-processed (non-spliced) RNA. Furthermore, this kit can be used to isolate RNA for downstream applications where it is necessary to avoid DNA contamination, since the cytoplasmic fraction has been shown to be free of all traces of genomic DNA.
RNA Yield HeLa (1 x 106) – Cytoplasmic RNA HeLa (1 x 106) – Nuclear RNA
15 µg ≤ 3.5 µg
Storage Conditions and Product Stability All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
Our Blyscan™ Glycosaminoglycan Kit has been a ‘go-to’ Solution for reliable sGAG and Proteoglycan Analysis for many years! Blyscan utilises a dye-binding approach to quantitatively measure sulfated glycosaminoglycans (sGAG) and proteoglycans in cells, tissues and fluids from a wide range of in-vivo and in-vitro sources.
Colorimetric Detection (656nm) (Endpoint)
Understanding Glycosaminoglycans (GAGs) and Proteoglycans
Glycosaminoglycans (GAGs) are a type of negatively charged polysaccharide that play crucial roles in various biological processes. They are composed of repeated disaccharide units, typically of N-acetylated or N-sulfated hexosamine paired with a uronic acid (GlcA or IdoA) or galactose. Sulfate groups can also be added to give sulfated GAGs an overall negative charge that influences cell interactions and also enable binding by our Blyscan dye reagent.
Common examples of GAGs include Chondroitin Sulfate, Dermatan Sulfate, Heparin, Heparan Sulfate, and Keratan Sulfate. Note that Hyaluronic Acid is a non-sulfated GAG and cannot be detected by the Blyscan assay. If you need to measure hyaluronic acid instead, we recommend using our Purple-Jelley kit!
The Role of Glycosaminoglycans in Tissues
GAGs and proteoglycans have essential functions in tissues and organisms, providing biophysical support through scaffolding and maintaining cartilage hydration. They also play a vital role in biochemical processes such as cell adhesion and signalling.
What is the origin of the Blyscan assay name?
Blyscan is an Old English word meaning ‘to shine’ and from which the word ‘blush’, (blushing), may have been derived. This was an appropriate choice as the Blyscan Assay contains a blue dye which ‘blushes’ bright pink when it binds to sulphated glycosaminoglycans!
How does the Blyscan assay work?
Step 1. Blyscan dye reagent contains DMMB dye in an optimised buffer. Addition of Dye reagent to samples containing sGAG results in the formation of a dye/sGAG complex due to a charge interaction between dye and GAG sulfate groups.
Step 2. Over a 30 minute incubation Dye-labelled sGAGs precipitate out of solution and are collected by centrifugation. Following removal of unbound dye, the remaining bound dye is released from the complex by addition of dye dissociation reagent. Released dye is quantified spectrophotometrically.
Step 3. The sGAG content of unknown samples may be quantified by comparison against a calibration curve prepared using a standard of purified Chondroitin-4-sulfate supplied with the kit.
A list of suggested sample types can be found under the ‘Assay Specification‘ tab.
The Blyscan Dye reagent is formulated to miminise binding to other charged sample components such as nucleic acids, a problem with some older dye-based sGAG assays.
Assay range
2.5 – 50µg/ml
Limit of Detection
2.5µg/ml
Detection Method
Colorimetric Detection (656nm) (Endpoint)
Measurements per kit
110 in total (allows a maximum of 48 samples to be run in duplicate alongside a standard curve).
In-vivo: Liquid samples, including fluids such as urine, amniotic or synovial fluid.
In-vitro: Solid samples, such as deposited ECM on 2D/3D culture surfaces.by enzymatic treatment
In-vivo: Liquid samples, Culture media during 2D/3D cell culture.
The assay requires that sulfated polysaccahrides or sGAGs are in a soluble form. A preliminary enzymatic extraction step is required for solid samples (enzyme not supplied with kit).
The assay is not suitable for use with samples containing alginates or that comprise degraded sulfated disaccharide fragments.
Precautions
This kit is designed for research use only. Not for use in diagnostic procedures. Kit requires access to a centrifuge, as well as a spectrophotometer/colorimeter capable of absorbance detection at 656nm. Specific sample preparation protocols may require customer to provide further reagents, consult assay manual for further information.
Blyscan sGAG kit contents:
1. Blyscan Dye Reagent (1x110ml)
2.sGAG Reference Standard (1x5ml, 100µg/ml Bovine tracheal chondroitin 4-sulfate)
3. Dissociation Reagent (1x110ml)
4. Sodium Nitrite (1x15ml)
5. Acetic Acid (1x15ml)
6. Ammonium Sulfamate (1x15ml)
7. 1.5ml micro-centrifuge tubes for dye-labelling reaction.
8. Assay kit manual
NB: Additional reagents may be required for sample preparation prior to assay. Consult manual or contact us for further details.
Document
Our Blyscan™ Glycosaminoglycan Kit has been a ‘go-to’ Solution for reliable sGAG and Proteoglycan Analysis for many years! Blyscan utilises a dye-binding approach to quantitatively measure sulfated glycosaminoglycans (sGAG) and proteoglycans in cells, tissues and fluids from a wide range of in-vivo and in-vitro sources.
Colorimetric Detection (656nm) (Endpoin
Rapid and efficient removal of abundant proteins from serum and plasma samples
Process 10 samples in 30 minutes
Allows for visualization of low abundance proteins
Convenient and affordable spin column protocol
Generic protocol allows for depletion from human and other animal samples
Purification is based on spin column chromatography that uses Norgen’s resin separation matrix
This kit provides a fast and simple procedure for the effective depletion of major serum proteins including albumin, α-antitrypsin, transferrin and haptoglobin from serum and plasma samples. Such abundant proteins usually obscure less abundant proteins in gel electrophresis making them difficult to visualize and recover. The kit reduces sample complexity to allow for convenient investigation of less abundant proteins. In other applications such as mass spectroscopy, fractionating abundant proteins improves the resolution of less abundant proteins. The kit is ‘generic’ in that it is not antibody-based and can thus be used to deplete abundant proteins from any starting material – human or various other animals – making it more versatile, flexible and affordable than other kits. Resulting proteins are ready for a range of applications including 2D gel electrophoresis, SDS-PAGE, Mass Spectrometry, Protein microarrays, DIGE (Difference Gel Electrophoresis) and more.
Storage Conditions All 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.
Short term stability: Ambient, Long term stability: See individual component labels
Stability:
> 2 years under recommended storage conditions
Analyte:
Pectin
Assay Format:
Spectrophotometer
Detection Method:
Absorbance
Wavelength (nm):
235
Signal Response:
Increase
Reaction Time (min):
~ 30 min
Application examples:
Food ingredients (e.g. citrus fruit and apple) and other materials.
The Pectin Identification Assay Kit is suitable for the identification of pectin in food ingredients. This kit now employs a new pectate lyase from Aspergillus niger.
All reagents stable for > 2 years after preparation
Only enzymatic kit available
Simple format
Standard included
Document
The Pectin Identification Assay Kit is suitable for the identification of pectin in food ingredients. This kit now employs a new pectate lyase from Aspergillus niger.