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多肽与蛋白> 蛋白检测> 蛋白定量
Bradford蛋白浓度测定试剂盒
产品编号: P0006
产品包装:1000次
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价格:¥116.00元
产品简介
使用说明
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产品编号 产品名称 产品包装 产品价格
P0006 Bradford蛋白浓度测定试剂盒 1000次 116.00元

碧云天生产的Bradford蛋白浓度测定试剂盒(Bradford Protein Assay Kit)是根据最常用的两种蛋白浓度检测方法之一
Bradford法研制而成,实现了蛋白浓度测定的快速、稳定和高灵敏度,并且检测数据有很好的线性关系。
Bradford法的检测原理是考马斯亮蓝(Coommassie Brilliant Blue)G-250与蛋白质的碱性和芳香族氨基酸特别是精氨酸
(Arginine)在酸性介质中结合后,溶液转变为蓝色,溶液最大吸收峰从465nm迁移到595nm,颜色的变化与蛋白质浓度成正比。因此,可通过检测595nm处的吸光度对溶液中蛋白质浓度进行测定。
本试剂盒检测速度极快,10-20个样品只需不足10分钟即可完成,和BCA法相比大大缩短了检测时间。
本试剂盒检测灵敏度高,最小蛋白检测量达到0.5μg。
本试剂盒标准曲线的线性关系好。常规的Bradford法的标准曲线是非线性的。碧云天的本试剂盒进行了精心优化,确保5μl体积的样品或标准品在0.1-1.5mg/ml浓度范围内有良好的线性关系(参考图1)。


图1. 本试剂盒蛋白标准曲线的实测效果图。图中数据仅供参考,实际的检测效果可能会略有不同。

与测定蛋白浓度的其它方法如BCA法和Lowry相比,Bradford法测定蛋白浓度不受绝大部分样品中的化学物质的影响,尤其是还原试剂的影响。样品中β-巯基乙醇(β-Mercaptoethanol)的浓度可高达1M,二硫苏糖醇(DTT)的浓度可高达5mM。但本试剂盒受略高浓度的去垢剂影响,需确保SDS低于0.01%,Triton X-100低于0.05%,Tween 20、60、80低于0.015%。含去垢剂的样品推荐使用碧云天生产的Bradford蛋白浓度测定试剂盒(去垢剂兼容型)(P0006C)或碧云天生产的BCA蛋白浓度测定试剂盒(P0009, P0010, P0010S, P0011, P0012, P0012S)。
Bradford蛋白浓度测定试剂盒对样品中各种物质的详细的兼容性,请参见碧云天如下网页:
http://www.beyotime.com/Compatibility Chart For Bradford Kit.pdf
      每个试剂盒可以检测1000个样品。
包装清单:

产品编号 产品名称 包装
P0006-1 G250染色液 125ml×2
P0006-2 蛋白标准(5mg/ml BSA) 1.5ml
说明书 1份

保存条件:
4℃保存,一年有效。蛋白标准可以-20℃长期保存。
注意事项:
蛋白标准请在全部溶解后先混匀,再稀释成一系列不同浓度的蛋白标准。
将G250染色液回复到室温再使用,有利于提高检测的灵敏度。
需可检测560-610nm之间波长的酶标仪一台,最佳检测波长为595nm。并需96孔板。
本产品仅限于专业人员的科学研究用,不得用于临床诊断或治疗,不得用于食品或药品,不得存放于普通住宅内。
为了您的安全和健康,请穿实验服并戴一次性手套操作。

使用说明:
1. 蛋白标准的准备

a. 蛋白样品在什么溶液中,标准品也宜用什么溶液稀释。但是为了简便起见,如果蛋白样品所在溶液不含有干扰本试剂盒检测的物质,也可以用0.9%NaCl、PBS或水稀释标准品。蛋白标准(5mg/ml BSA)如果冻存,请完全融化并混匀后使用。
b. 按照下表配制0、0.125、0.25、0.5、0.75、1、1.5mg/ml蛋白标准。每次稀释时注意充分混匀。如果有必要可以增加设置0.0625mg/ml的蛋白标准。

编号 稀释液体积 标准品体积 最终浓度
A 70μl 5mg/ml BSA 30μl 1.5mg/ml
B 30μl 从A管取60μl 1mg/ml
C 20μl 从B管取60μl 0.75mg/ml
D 30μl 从C管取60μl 0.5mg/ml
E 60μl 从D管取60μl 0.25mg/ml
F 60μl 从E管取60μl 0.125mg/ml
G 60μl 0μl 0mg/ml

2. 蛋白浓度测定
a. 取5μl不同浓度蛋白标准加到96孔板的蛋白标准孔中。
b. 取5μl样品到96孔板的样品孔中。如果样品不足5μl,需加标准品稀释液补足到5μl。请注意记录样品体积。
c. 各孔加入250μl G250染色液
d. 用酶标仪测定A595,或560-610nm之间的其它波长的吸光度。可以立即测定吸光度,也可以在2小时内测定,2小时内检测数据无显著变化。
e. 根据标准曲线和使用的样品体积计算出样品中的蛋白浓度。

相关产品:

产品编号 产品名称 包装
P0006 Bradford蛋白浓度测定试剂盒 1000次
P0006C Braford蛋白浓度测定试剂盒(去垢剂兼容型) 800次
P0007 蛋白标准(5mg/ml BSA) 1ml
P0009 BCA蛋白浓度测定试剂盒(增强型) 5000次
P0010 BCA蛋白浓度测定试剂盒(增强型) 500次
P0010S BCA蛋白浓度测定试剂盒(增强型) 200次
P0011 BCA蛋白浓度测定试剂盒 5000次
P0012 BCA蛋白浓度测定试剂盒 500次
P0012S BCA蛋白浓度测定试剂盒 200次
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