各位,请教一下,购买了Sigma的硫酸软骨素酶ABC,准备做动物实验用的,但是瓶子里是冻干粉剂,而且是5UI量(Specific Activity: 50–250 units/mg protein (using chondroitin sulfate C as substrate)),使用说明是这样描述的:Preparation Instructions :Reconstitute in a 0.01% bovine serum albumin aqueous solution. Subsequent dilutions can be made into a buffer containing 50 mM Tris, pH 8.0, 60 mM sodium acetate and 0.02% bovine serum albumin. Solutions should be prepared fresh.准备说明:0.01%牛血清白蛋白水溶液复溶。随后的稀释液应该现配现用,可以被制成含有50mM Tris,pH值8.0,60 mM乙酸钠和0.02%牛血清白蛋白的溶液。
有几个问题请教:第一,0.01%BSA溶液加多少进入干粉内较适宜?第二,50mM Tris 是怎么配制的?第三,最后的含三种成分比例在配制过程中如何把握?我需要的终浓度含ChABC为1UI/ml,又该怎么配制?