Bicolor’s Purple-Jelley Assay（透明質(zhì)酸檢測試劑盒）采用Stains-all染料結(jié)合法定量檢測哺乳動物組織中的透明質(zhì)酸。該方法詳細(xì)描述了在測定分離的透明質(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

H1000透明質(zhì)酸檢測試劑盒對樣本的要求：皮膚、軟骨、軟組織，包含不低于0.2ug的透明質(zhì)酸。

Bicolor’s Purple-Jelley Assay（透明質(zhì)酸檢測試劑盒）實(shí)驗(yàn)操作步驟：

蛋白的去除：所有測試樣本都需要進(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).