Blyscan™ – sulfated Glycosaminoglycan (sGAG) assay kit

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).

Suitable Samples

In-vivo: Solid samples, including cartilage, bone, connective tissue, tumour tissue

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.

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

Introduction

Magen’s HiPure columns are prepared by high quality glass fiber filter membrane as raw materials through membrane cutting, membrane release, ring release, ring pressing, gland, weighing and other processes. HiPure nucleic acid adsorption columns have the characteristics of long-term stability and high binding capacity. Experiments show that the highest binding capacity and binding efficiency of HiPure nucleic acid adsorption columns are basically unchanged when stored at room temperature for 4 years.

The series of nucleic acid columns produced by Magen Biotech are based on carefully selected imported glass fiber membranes (GF/B, GF/D, GF/F). Columns production processes such as polypropylene injection molding materials, injection molding process, and downstream membrane packing and compression rings are strictly controlled. This is to ensure that the column has extremely high adsorption capacity and long-term stability. Compared with conventional products on the market, Magen’s columns are with varieties, and binding rate will not change when stored at room temperature for 4 years.

Details

Specifications

Features	Specifications
Recommended application	gDNA and RNA Isolation
Preservation conditions	Room temperature
Stability	Up to 4 years
Filter membrane	High quality glass fiber filter GF/B, 2 layers
Membrane aperture	1.0μm
Maximum binding yield of plasmid	30 μg
Maximum yield of alcohol mediated Binding	100 μg
Single liquid carrying capacity of column	900 μl
Minimum elution volume	80 μl
Withstand centrifugal force	4,000-5,000 x g
Centrifuge	Low speed centrifuge, Swing out Rotor, can placed a height of 6.5cm square,  (height of HiPure DNA Plate & 1.6ml Collection Plate: height, 6.2cm)

Adsorption Mechanism

Based on the negatively charged DNA skeleton, it has a high affinity for positively charged glass fibers. In high salt and ethanol solutions, DNA/RNA binds to glass fiber and interacts with hydrophilic matrix on silica through hydrogen bond. DNA/RNA is tightly bound. All pollutants can be removed by washing solution. At high salt concentration, nucleic acids selectively bind to silica gel membrane, while other pollutants, mainly proteins, are removed by membrane washing.

Ordering information

CAT.No.	Product Name	Package
C13131	HiPure gDNA Plate (2 x GF/B)with 1.6ml Collection Plate	10/Bag

Purchase Guide

Item No.	Product Name	Membrane type/number of layers	Collection tubes	Plasmid DNA binding capacity (Physical adsorption)	gDNA/RNA binding capacity (Alcohol-mediated adsorption)	Minimum Elution volume	Liquid volume capacity
C13010	HiPure DNA Nano Column	2 layers GF/F	2ml without cap	5μg	20μg	10μl	700μl
C13011	HiPure DNA Micro Column	3 layers GF/F	2ml without cap	10μg	50μg	15μl	700μl
C13100	HiPure DNA Mini Column I	2 layers GF/B	2ml without cap	15μg	100μg	30μl	700μl
C13110	HiPure DNA Mini Column II	4 layers GF/B	2ml without cap	35μg	200μg	50μl	800μl
C13111	HiPure RNA Mini Column	3 layers GF/B	2ml without cap	30μg	200μg	30μl	800μl
C13112	HiPure Viral Mini Column	3 layers GF/F	2ml without cap	30μg	200μg	30μl	800μl
C13113	HiPure CFDNA Mini Column	3 layers GF/F,1 layer GF/B	2ml without cap	30μg	200μg	30μl	800μl
C13120	HiPure DNA Midi Column	4 layers GF/B	15ml Centrifuge tube	125μg	1mg	500μl	4ml
C13121	HiPure DNA Midi Column III	8 layers GF/B	15ml Centrifuge tube	250μg	1mg	500μl	4ml
C13122	HiPure DNA Maxi Column	4 layers GF/B	50ml Centrifuge tube	500μg	5mg	1000μl	20ml
C13123	HiPure DNA Maxi Column III	8 layers GF/B	50ml Centrifuge tube	1mg	5mg	1000μl	20ml
C13124	HiPure DNA Maxi Column C	8 layers GF/B	50ml high speed Centrifuge tube	1mg	5mg	700μl	12ml
C13130	HiPure DNA Plate	2 layers GF/F	1.6ml Plate	30μg	100μg	80μl	900μl
C13131	HiPure gDNA Plate	2 layers GF/B	1.6ml Plate	30μg	100μg	80μl	900μl

Note: GF/B pore size is for 1.0μM glass fiber membrane; GF/F pore size is for 0.7μm glass fiber membrane.

Magen’s HiPure columns are prepared by high quality glass fiber filter membrane as raw materials through membrane cutting, membrane release, ring release, ring pressing, gland, weighing and other processes. HiPure nucleic acid adsorption columns have the characteristics of long-term stability and high binding capacity. Experiments show that the highest binding capacity and binding efficiency of HiPure nucleic acid adsorption columns are basically unchanged when stored at room temperature for 4 years.

Description

Specifications

Clone	IHC580
Source	Mouse Monoclonal
Positive Control	Pituitary
Dilution Range	1:200

Follicle-Stimulating Hormone (FSH) allows for progression of ovarian folliculogenesis, and enables Sertoli cell proliferation in the testis. Anti-FSH reacts with FSH-producing cells, therefore FSH staining is useful for classifying pituitary cancers and understanding pituitary disease.