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.

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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.

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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

Product Description

    Product Detail

Applicable equipment:

It is recommended to use the Isothermal fluorescence detector developed by Amp-future, which is also suitable for fluorescence quantitative PCR apparatus with market known brands.

Kit Storage and Term of Validity

Storage term: stored at ≤-20℃,keep away from light, avoid heavy weight and repeated freezing and thawing.

Term of Validity: 14 months

Isothermal Nucleic Acid Principle Summary

This kit is based on a rapid nucleic acid amplification technology at room temperature and constant temperature: at room temperature and constant temperature (generally 39℃~42℃), with the help of auxiliary proteins and single-strand binding proteins,the recombinase and primers form a complex;Source search and combine the target homology domain, at this time,a D-loop region is formed at the homology position and strand exchange begins;along with the dissociation of the recombinase from the complex,the polymerase also binds to the 3′ end of the primer and begins chain extension. At the same time, relying on the function of exonuclease, adding specific molecular probes designed according to the template, and using fluorescence monitoring equipment can realize real-time monitoring of the amplification process of the target fragment.

Isothermal Nucleic Acid Product Features

1/ High sensitivity and specificity, short reaction time.

2/ The reagent form is freeze-dried, stable and easy to operate.

Technical Parameters:

Parameters	Details
Product Name	DNA Isothermal Amplification Kit EXO
Manufacturer	Amp-future
Storage Temperature	-20°C
Kit Components	Enzymes, Buffers ,Reagents
Packaging	48 Tests/box
Detection Limit	500-1000copies/µL
Shipping	ICE
Test Time	5-20mins

Isothermal Nucleic Acid Applications

Suitable for DNA isothermal rapid amplification kit(fluorescent type)

Primer: Require pair of nucleotide primers with the length of 25-35 bp.

Fluorescent Probe:Require the suitable length is 46-52nt.

DNA fluorescent kit reaction temperature is 39 to 42℃ and time is 5-20 minutes.

Notes

1/ Please avoid nucleic acid contamination and set blank control during reaction due to the high sensitivity of the kit.

2/ Please take out the required quantity of MIRA reaction units for the experiment, and put the rest under storage conditions when performing the experiment.