The amount of glycosaminoglycans (GAGs) in the lyophilized tissue sample of normal and decellularized esophagi (n = 7) were measured using the Blyscan Sulfated Glycosaminoglycan Kit (Biocolor). The tissue samples were digested with papain extraction reagent provided with the kit at 65°C. The extracted GAGs were then incubated with Blyscan dye reagent to form a precipitate of sulfated GAG dye complex. This precipitate was further dissociated by the dissociation reagent provided with the kit and absorbance was measured at 656 nm on a plate reader.
Blyscan sulfated glycosaminoglycan kit
Manufactured by Biocolor
Sourced in United Kingdom
The Blyscan sulfated glycosaminoglycan kit is a laboratory assay used to quantify the amount of sulfated glycosaminoglycans present in a sample. It provides a colorimetric method for the detection and measurement of these complex carbohydrates.
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3 protocols using blyscan sulfated glycosaminoglycan kit
1
Quantification of GAGs in Esophageal Tissue
2
GAG Quantification in Brain and Liver
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Brain samples and liver samples were lysed in water by Tissue Lyser equipment. The lysates were then digested with Proteinase K, and extracts were clarified by centrifugation and filtration. GAG levels in brain extracts were determined using the Blyscan sulfated glycosaminoglycan kit (Biocolor, Carrickfergus, UK) with chondroitin 4-sulfate as the standard.
3
Glycosaminoglycan Quantification in Tissues
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Tissues were weighed and digested overnight in 0.2 mg/ml proteinase K solution. Extracts were centrifuged and supernatants were clarified by filtration in 0.22 μm microporous membrane-containing filters (Ultrafree MC, Millipore). GAG content was determined using the Blyscan sulfated glycosaminoglycan kit (Biocolor) using chondroitin 4-sulfate as standard and was normalized to wet tissue weight. Owing to the inherent technical variability of the assay, the GAG content of WT animals at each age was set to 1 to allow comparison of data obtained at different points in time.
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