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产品介绍
Bicolor’s Purple-Jelley Assay(H1000透明质酸检测试剂盒)采用Stains-all染料结合法定量检测哺乳动物组织中的透明质酸。该方法详细描述了在测定分离的透明质酸之前,通过两步关键的电解质盐析去除组织中蛋白和硫酸化糖胺聚糖的过程。
部分组织中透明质酸的分布情况:
Distribution of HA in Tissue
Species | Tissue | Concentration (µg/g) |
Mouse | heart | 48 |
kidney | 11 | |
lung | 14 | |
leg muscle | 20 | |
skin | 307 | |
Rat | heart | 83 |
kidney | 28 | |
lung | 17 | |
leg muscle | 67 | |
skin | 510 | |
Rabbit | heart | 138 |
kidney | 27 | |
lung | 45 | |
leg muscle | 50 | |
skin | 428 |
H1000透明质酸检测试剂盒对样本的要求:皮肤、软骨、软组织,包含不低于0.2ug的透明质酸。
H1000透明质酸检测试剂盒实验操作步骤:
蛋白的去除:所有测试样本都需要进行蛋白质消化。
糖胺聚糖的复苏:通过GAG Precipitation Reagent 获得释糖胺聚糖。
透明质酸的分离测定:离心洗涤获得透明质酸水合物后,绘制标准曲线,655nm处定量检测透明质酸。
产品订购:
Standard Assay Kit: Product Code H1000 (100 assays)
Economy Assay Kit: Product Code H2000 (400 assays)
参考文献
1.Chemistry and Biology of Hyaluronan, Edited by H.G. Garg and C.A. Hales, (2004). Publisher: Elsevier Oxford, UK
2.Hyaluronan in Cancer Biology, Edited by R.Stern, (2009) Publisher: Academic Press, San Diego, U.S.A
3.In 1936 Edwin Jelley sent a ‘Letter to the Editor’ of Nature, (138 1009 -1010), regarding the unusual behaviour of some cyanine dyes.
4.When dissolved in 5 M NaCl these dyes produce a third absorbance peak at a longer wave-length (650nm) whereas in deionised water only a double peak occurs at ~540 nm and ~570 nm.
5.The 650 nm peak in concentration dye solution induces aggregation of the dye molecules and has been described as a ‘J-aggregate’ named after Jelley.
6.Individual dye molecules may form a stacking format to produce a supra-
molecular complex as seen in peak 3, (Image on front cover of the Manual)
7.Further studies in the 1960s notably by Kay et.al. (J. Physical Chem. 68 1896 – 1906) found many biological polymers including proteins, DNA, polar lipids and glycosaminoglycans could induce the third peak when using high dye concentrations.
8.The dye was renamed ‘Stains-all’ by Dhlberg, Dingman and Peacock in1969
( J. Mol. Biol. 41, 139).