Glycosaminoglycans (GAGs) are negatively-charged polysaccharides composed of repeated disaccharide units. These units typically consist of N-acetylated or N-sulfated hexosamine along with a uronic acid (GlcA or IdoA) or galactose. Sulfate groups may also be added, influencing cell interactions within tissues. Common examples of GAGs include Chondroitin Sulfate, Dermatan Sulfate, Heparin, Heparan Sulfate, and Keratan Sulfate. Notably,Hyaluronic Acid is a non-sulfated GAG (which can be specifically detected and measured using our Purple-Jelley kit!).
When one or more GAG chains are covalently linked to a core protein, the resulting molecule is referred to as a proteoglycan (PG).
Glycosaminoglycans (GAGs) and proteoglycans have a wide range of functions in tissues and organisms. They provide essential biophysical support, including acting as scaffolds and maintaining cartilage hydration. Additionally, they play a vital role in biochemical processes, including cell adhesion and cell signalling.
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!
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.
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.
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.
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.
Experience user-friendly detection & measurement of Soluble Collagen with Sircol™ 2.0! Our latest kit simplifies collagen quantification within in-vivo / in-vitro samples. Sircol 2.0 offers enhanced sensitivity and accuracy compared to our previous Sircol kit.
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.
Biocolor’s Purple-Jelley assay kit is the perfect tool for accurate measurement of hyaluronic acid / Hyaluronan levels in your samples. This colorimetric assay is optimised for quantitative analysis in-vivo, tissue-derived hyaluronic acid / Hyaluronan and includes full step-by-step instructions.
