Bicolor’s Purple-Jelley Assay(H1000透明質(zhì)酸檢測(cè)試劑盒)采用Stains-all染料結(jié)合法定量檢測(cè)哺乳動(dòng)物組織中的透明質(zhì)酸。該方法詳細(xì)描述了在測(cè)定分離的透明質(zhì)酸之前,通過兩步關(guān)鍵的電解質(zhì)鹽析去除組織中蛋白和硫酸化糖胺聚糖的過程。
部分組織中透明質(zhì)酸的分布情況:
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透明質(zhì)酸檢測(cè)試劑盒對(duì)樣本的要求:皮膚、軟骨、軟組織,包含不低于0.2ug的透明質(zhì)酸。
H1000透明質(zhì)酸檢測(cè)試劑盒實(shí)驗(yàn)操作步驟:
蛋白的去除:所有測(cè)試樣本都需要進(jìn)行蛋白質(zhì)消化。
糖胺聚糖的復(fù)蘇:通過GAG Precipitation Reagent 獲得釋糖胺聚糖。
透明質(zhì)酸的分離測(cè)定:離心洗滌獲得透明質(zhì)酸水合物后,繪制標(biāo)準(zhǔn)曲線,655nm處定量檢測(cè)透明質(zhì)酸。
產(chǎn)品訂購:
Standard Assay Kit: Product Code H1000 (100 assays)
Economy Assay Kit: Product Code H2000 (400 assays)
參考文獻(xiàn)
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).