Blyscan™ GLYCOSAMINOGLYCAN Assay

Assay time: 1 hour     Assay sensitivity: 0.25µg

Blyscan Glycosaminoglycan Kit FAQs

The Blyscan™ Assay is a quantitative dye-binding method for the analysis of sulfated proteoglycans and glycosaminoglycans.

Sulfated glycosaminoglycan (sGAG) forms that can be measured; soluble salt extracts from:

1. in vitro studies where extracellular matrix components are released by live cells into the cell culture medium;
2. elastic, fibrous and hyaline cartilages;
3. arteries, lung, skin and other material containing extracellular matrix (connective tissue);
4. solid tumour specimens;
5. cell and tissue extracts;
6. aliquots from protein chromatography fractions
7. sulfated glycosaminoglycans in amniotic fluid and urine samples can be measured;
8. the assay may be adopted to detect and/or measure sulfated glycosaminoglycan degradative enzymes, and;
9. the N- and O-sulfated glycosaminoglycan ratio can be measured

Blyscan Updated Manual Blyscan  Citations

Blyscan Method Summary

1. Test materials, standards and reagent blanks (100μl) are mixed with Dye Reagent (1ml) in microcentrifuge tubes for 30 minutes. The glycosaminoglycan-dye complex formed precipitates out of solution.
2. The complex is packed firmly to the bottom of the tubes by centrifugation at 12,000g for 10 minutes.
3. Unbound dye is removed by inverting and draining tubes.
4. Bound dye is released and dissolved via addition of Dissociation Reagent (1ml) and vortex mixing for 5-15 min.
5. Released dye is measured spectrophotometrically at 656nm or colorimetrically, usually with a red filter, using a multiwell plate reader.

The pictures below outline the Blyscan™ Assay:

	[1] Reagent blank (left) and Reference standard (right), mixed with 1 ml Blyscan Dye reagent Note: Pink ‘blush’ in the Reference standard tube [2] Tubes centrifuged and supernatant removed Note: Purple pellet of dye-sGAG complex in Reference standard tube [3] Tubes after addition of the Blyscan Dye Dissociation reagent Note: The pink/purple colour of the dye-sGAG complex has been replaced by the blue colour of the “free” dye

Blyscan Kit Components

• Dye Reagent: 1,9-dimethyl-methylene blue in an organic buffer containing surfactants and stabilisers

• Dissociation Reagent: a chaotropic salt in an aqueous propan-1-ol solution

• Sulfated Glycosaminoglycan Standard: bovine chondroitin 4-sulfate. Concentration: 100μg/ml

• Sodium Nitrite Solution: 5% (w/v)

• Acetic Acid Solution: 33% (v/v)

• Ammonium Sulfamate Solution: 12.5% (w/v)

Blyscan™ Sulfated Glycosaminoglycan Products

Standard Assay Kit (120 assays) Product Code: B1000

Large Economy Assay Kit (475 assays) Product Code: B3000

Dye Reagent & Dissociation Reagent Pack (2 x 120ml Blyscan Dye Reagent, plus 2 x 120ml Blyscan Dissociation Reagent) Product Code: B1500

Dye Reagent Pack (3 x 120ml) Product Code: B1005

Glycosaminoglycan Standard (100μg/ml; 3 x 5ml) Product Code B1010

Glycosaminoglycan Isolation & Concentration Pack to clean up and concentrate samples (sufficient for 100 samples) Product Code B1015

Order

Blyscan Glycosaminoglycan Kit FAQs