Biocolor 可溶性胶原检测试剂盒BiocolorSircol Soluble Collagen Assay采用与胶原特异性结合的Sircol染料,主要用于定量检测哺乳动物组织中抽提的酸溶性胶原和体外培养哺乳动物细胞释放到培养基中的胶原。溶于酸、胃蛋白等的可溶性胶原均可检测,具体可检测的胶原类型为I-V型,VI-XIV型胶原也可检测,但由于难以制备标准曲线,还无法定量。Sircol染料不能区分不同类型的胶原,但可与胶原的[Gly-X-Y]n螺旋结构特异性地结合。随着胶原变性为凝胶,该染料与胶原的结合力减弱。非脊椎动物与胶原由于变性温度较低、羟脯氨酸残基较少等原因,结合染料的能力较弱,需要制备种属特异性的纯化标准品,以绘制标准曲线。实验步骤
绘制标准曲线 待测样本的制备:取50ul样本容易调整体积到100ul进行反应。根据反应的结果对样本浓度进行调整,使其在标准曲线范围内; 胶原 染料复合物的获得:加入1ml Sircol染料,混匀离心后,既得胶原 染料复合物,弃上清,去掉未结合的染料 加入1ml 染料释放剂,将胶原 染料复合物中的染料释放出来 吸光度测定:用分光光度计540nm出定量测定。或使用colorimeter测定产品订购Product Code: S1000120 assaysProduct Code: S5000475 assays参考文献1Anilkumar, 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.2Arizmendi, N., Puttagunta, L., Chung, K. L., Davidson, C., Rey-Parra, J., Chao, D. V., Thebaud, B., Lacy, P. andVliagoftis, H. (2014) Rac2 is involved in bleomycin-induced lung inflammation leading to pulmonary fibrosis. RespiratoryResearch 15, 71.3Aschner, Y., Khalifah, A. P., Briones, N., Yamashita, C., Dolgonos, L., Young, S. K., Campbell, M. N., Riches, D. W.,Redente, E. F., Janssen, W. J., et al. 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