蛋白质定量试剂盒 (BCA法) P1511
货号 | 产品名称 | 规格 | 价格 |
P1511-1 | 蛋白定量试剂盒 (BCA法) | 500次 | 330元 |
P1511-2 | 蛋白定量试剂盒 (BCA法) | 2500次 | 1300元 |
P1511-3 | 蛋白定量试剂盒 (BCA法) | 200次 | 180元 |
描述:Bicinchoninic acid (BCA )法是近来广为应用的蛋白定量方法。其原理与Lowery法蛋白定量相似,即在碱性环境下蛋白质与Cu2+络合并将Cu2+还原成Cu1+。BCA与Cu1+结合形成稳定的紫蓝色复合物,在562 nm处有高的光吸收值并与蛋白质浓度成正比,据此可测定蛋白质浓度。与Lowery法相比,BCA蛋白测定方法灵敏度高,操作简单,试剂及其形成的紫兰色复合物稳定性俱佳,并且受干扰物质影响小。与Bradford法相比,BCA法的显著优点是不受去垢剂的影响。
特点:
1、 与Lowery法相比,BCA蛋白测定方法灵敏度高,操作简单,试剂及其形成的紫兰色复合物稳定性俱佳,并且受干扰物质影响小;
2、 与Bradford法相比,BCA法的显著优点是不受去垢剂的影响。
规格:500次 2500次 200次
储存:(1) BCA Reagent 100 mL,室温保存;(2) Cu Reagent 2 .5mL,室温保存;(3) BSA standard 4 mg/mL 1 mL,−20ºC冻存。12个月有效
操作步骤:
1、工作溶液(Working Reagent, WR)配制
2、标准蛋白溶液配制
3、蛋白浓度测定和数据分析
注意事项:
1. 37ºC 30min或25ºC室温反应2小时对测量较为便利,但严格来讲此时反应尚未达到终点,通常每10 min OD562值升高约2.3%。然而,通常在10min内可以测定30管并不会明显影响测定精度。
2. BCA法检测范围为20-2000 µg/mL。检测0.5~10 µg/mL高度稀释蛋白样品应采用Bradford法蛋白质定量试剂盒(P1510)。60ºC 30 min反应可增加检测灵敏度至5~250µg/mL。
3. BCA法检测样品中含有脂类物质时光吸收值会偏高。样品中EDTA或葡萄糖浓度大于10 mM不能使用BCA方法,葡萄糖浓度大于10 mM时可用改良Lowry法蛋白定量试剂盒 #P1512,EDTA大于10 mM的样品可用Bradford法蛋白质定量试剂盒(P1510)。另外,蛋白质样品经液体样品蛋白抽提试剂(P1255)沉淀后,可彻底去除干扰BCA法、Bradford法、和Lowry法蛋白测定的物质。
4. 欲使测量能耐受下面表2所提示的最大干扰物质浓度,并保持测量精度,应在蛋白标准管中加入相应浓度的干扰物质,但会给操作带来不便。
5. 可测量吸附于固相支持物乳酶标板、琼脂糖、亲和层系凝胶上的蛋白。
6. 每次测定应该重新测定并制作标准曲线。
目前,使用普利莱BCA蛋白定量试剂盒发表的SCI文章已超过百篇,部分文章如下,供参考。
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