茜素红染色后定量 求助

daxi-fighting@1

我对我的干细胞进行了矿化诱导，做了茜素红染色，看到有的文献里 讲钙结节用10% cetylpyridinium chloride (CPC) in 10 mmol/L sodium phosphate 37°溶解半小时，后测562nm波长的吸光值。最终钙的水平还要以提取的细胞的全蛋白浓度来统一。组里至今没有做过钙浓度定量。所以请教一下大家。原文是这样说的At day 14, Alizarin Red staining was carried out and images were acquired using a scanner. Then, alizarin red was destained with 10% cetylpyridinium chloride (CPC) in 10 mmol/L sodium phosphate for 30 min at room temperature. The calcium concentrations were determined according to the absorbance at 562 nm using a standard calcium curve prepared in the same solution. The final calcium levels in each group were normalized with the total protein concentrations obtained from the duplicate plates. 现在我想请教问题：1：这个cpc如何配置？2：为什么还要乘以全蛋白浓度呢？3:标准品如何配置？标准曲线如何做？4：做这种ALP活性检测，还有钙浓度的检测，都必须乘以全蛋白浓度？不乘的话 如何？

robinpen

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; |1 ~% y5 m2 o: ^4 u. V应该是除以总蛋白浓度吧，相当于单位含量的蛋白中含有多少ALP或者Ca