Biocolor 硫酸氨基聚糖檢測試劑盒
Biocolor Blyscan™ Glycosaminoglycan Assay
采用常規生化方法,通過(guò)标準曲線和比色,可定量分析總硫酸化多糖(氨基聚糖和蛋白聚糖)的含量,亦可測定O端和N端硫酸氨基多糖的比值。主要采用的是Blyscan染料(Biocolor公司的專利産品),它可以特異性的與氨基聚糖和蛋白聚糖上的硫酸多糖組分結合。該方法不适于檢測已經(jīng)降解的硫酸化雙糖片段、
該試劑盒可檢測廣泛的蛋白多糖,可檢測的氨基多糖包括硫酸軟骨素、硫酸角質素、硫酸皮膚素、硫酸肝素,不适合檢測包含海藻酸的樣(yàng)品,因爲它們含有糖醛酸。該試劑盒也不能(néng)對(duì)透明質酸定量,但透明質酸并不幹擾對(duì)硫酸化氨基多糖的檢測。細胞培養基中20%的胎牛血清也不會(huì)幹擾實驗。該方法檢測靈敏度爲5ug,所需時(shí)間爲1小時(shí)
檢測樣(yàng)本:可來自軟骨組織、動脈組織、肺組織、皮膚組織、腫瘤組織、細胞核組織提取物、細胞培養液、羊水和尿液
實驗操作
1. 繪制标準曲線
2. 待測樣(yàng)品的處理:
取50ul待測樣(yàng)品進(jìn)行實驗。如果反應物在656nm初的吸收峰大于1.5,則對(duì)樣(yàng)品進(jìn)行稀釋,待測樣(yàng)樣(yàng)品濃度落入标準曲線中段時(shí),準确度較高。
3. 硫酸氨基聚糖—染料複合物的獲得
加1.0ml的Blyscan染料溶液到待測樣(yàng)本中,充分混勻,待不溶物出現,離心,棄上清,則獲得紫色的硫酸氨基聚糖—染料複合物
4. 硫酸氨基聚糖—染料複合物中染料的釋放
加入1.0ml的染料釋放劑,充分混勻。溶解5min以上,使染料充分釋放
5. 測定
用分光光度計于波長(cháng)656nm處測定,或使用colorimeter紅色濾光片測定
産品訂購
Product Code: B1000 110 assays
Product Code: B3000 440 assays
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