Biocolor 硫酸氨基聚糖检测试剂盒Biocolor Blyscan™ Glycosaminoglycan Assay 采用常规生化方法，通过标准曲线和比色，可定量分析总硫酸化多糖（氨基聚糖和蛋白聚糖）的含量，亦可测定O端和N端硫酸氨基多糖的比值。主要采用的是Blyscan染料（Biocolor公司的产品），它可以特异性的与氨基聚糖和蛋白聚糖上的硫酸多糖组分结合。该方法不适于检测已经降解的硫酸化双糖片段。 该试剂盒可检测广泛的蛋白多糖，可检测的氨基多糖包括硫酸软骨素、硫酸角质素、硫酸皮肤素、硫酸肝素，不适合检测包含海藻酸的样品，因为它们含有糖醛酸。该试剂盒也不能对透明质酸定量，但透明质酸并不干扰对硫酸化氨基多糖的检测。细胞培养基中20%的胎牛血清也不会干扰实验。该方法检测灵敏度为5ug，所需时间为1小时。检测样本：可来自软骨组织、动脉组织、肺组织、皮肤组织、肿瘤组织、细胞核组织提取物、细胞培养液、羊水和尿液实验操作1. 绘制标准曲线2. 待测样品的处理：取50ul待测样品进行实验。如果反应物在656nm初的吸收峰大于1.5，则对样品进行稀释，待测样样品浓度落入标准曲线中段时，准确度较高。3. 硫酸氨基聚糖 染料复合物的获得加1.0ml的Blyscan染料溶液到待测样本中，充分混匀，待不溶物出现，离心，弃上清，则获得紫色的硫酸氨基聚糖 染料复合物4. 硫酸氨基聚糖 染料复合物中染料的释放加入1.0ml的染料释放剂，充分混匀。溶解5min以上，使染料充分释放5. 测定用分光光度计于波长656nm处测定，或使用colorimeter红色滤光片测定产品订购Product Code: B1000120 assaysProduct Code: B3000475 assays参考文献1Almeida, H. V., Liu, Y., Cunniffe, G. M., Mulhall, K. J., Matsiko, A., Buckley, C. T., O Brien, F. J. and Kelly, D. J.(2014) Controlled release of TGF- 3 from cartilage extra cellular matrix derived scaffolds to promote chondrogenesis ofhuman joint tissue derived stem cells. Acta Biomaterialia.2Amin, H. D., Brady, M. A., St-Pierre, J.-P., Stevens, M. M., Overby, D. R. and Ethier, C. R. (2014) Stimulation ofChondrogenic Differentiation of Adult Human Bone Marrow-Derived Stromal Cells by a Moderate-Strength StaticMagnetic Field. Tissue Engineering Part A.3Anilkumar, T. V., Vineetha, V. P., Revi, D., Muhamed, J. and Rajan, A. (2014) Biomaterial properties of cholecystderived scaffold recovered by a non-detergent/enzymatic method. Journal of Biomedical Materials Research Part B:Applied Biomaterials.4Baiguera, S., Del Gaudio, C., Kuevda, E., Gonfiotti, A., Bianco, A. and Macchiarini, P. (2014) Dynamic decellularizationand cross-linking of rat tracheal matrix. Biomaterials 35, 6344 6350.5Baiguera, S., Del Gaudio, C., Lucali, E., Kuevda, E., Boieri, M., Mazzanti, B., Bianco, A. and Macchiarini, P. (2014)Electrospun gelatin scaffolds incorporating rat decellularized brain extracellular matrix for neural tissue engineering.Biomaterials 35, 1205 1214.6Bhattacharyya, S., Feferman, L. and Tobacman, J. K. (2014) Increased Expression of Colonic Wnt9A through Sp1-Mediated Transcriptional Effects involving Arylsulfatase B, Chondroitin-4-Sulfate, and Galectin-3. Journal of BiologicalChemistry jbc M114.7BUCKLEY, C. T. (2014) Shape-memory porous alginate scaffolds for regeneration of the annulus fibrosus-Effect of TGF-?3 supplementation and oxygen culture conditions.8Craciunescu, O., Gaspar, A., Trif, M., Moisei, M., Oancea, A., Moldovan, L. and Zarnescu, O. (2014) Preparation andCharacterization of a Collagen-Liposome-Chondroitin Sulfate Matrix with Potential Application for InflammatoryDisorders Treatment. Journal of Nanomaterials 2014.9Del Rey, M. J., Far , R., Izquierdo, E., Usategui, A., Rodr guez-Fern ndez, J. L., Su rez-Fueyo, A., Ca ete, J. D. andPablos, J. L. (2014) Clinicopathological Correlations of Podoplanin (gp38) Expression in Rheumatoid Synovium and ItsPotential Contribution to Fibroblast Plaet Crosstalk. PloS one 9, e99607.10Diederichs, S. and Tuan, R. S. (2014) Functional Comparison of Human-Induced Pluripotent Stem Cell-DerivedMesenchymal Cells and Bone Marrow-Derived Mesenchymal Stromal Cells from the Same Donor. Stem cells anddevelopment.11Faulk, D. M., Carruthers, C. A., Warner, H. J., Kramer, C. R., Reing, J. E., Zhang, L., D Amore, A. and Badylak, S. F.(2014) The effect of detergents on the basement membrane complex of a biologic scaffold material. Acta biomaterialia10, 183 193.12 Feferman, L., Bhattacharyya, S., Birch, L., Prins, G. S. and Tobacman, J. K. (2014) Differential effects of estrogenexposure on arylsulfatase B, galactose-6-sulfatase, and steroid sulfatase in rat prostate development. The Journal ofsteroid biochemistry and molecular biology 143, 105 114.13Gilpin, S. E., Guyette, J. P., Gonzalez, G., Ren, X., Asara, J. M., Mathisen, D. J., Vacanti, J. P. and Ott, H. C. (2014)Perfusion decellularization of human and porcine lungs: Bringing the matrix to clinical scale. The Journal of Heart andLung Transplantation 33, 298 308.14Gomez-Sanchez, C., Kowalczyk, T., Ruiz De Eguino, G., Lopez-Arraiza, A., Infante, A., Rodriguez, C. I., Kowalewski, T.A., Sarrionandia, M. and Aurrekoetxea, J. (2014) Electrospinning of poly (lactic acid)/polyhedral oligomericsilsesquioxane nanocomposites and their potential in chondrogenic tissue regeneration. Journal of BiomaterialsScience, Polymer Edition 1 24.