碧云天生物技术-Bradford蛋白浓度测定试剂盒(去垢剂兼容型)(P0006C)

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Bradford蛋白浓度测定试剂盒(去垢剂兼容型)

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产品编号: P0006C

产品包装:800次

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产品编号	产品名称	产品包装	产品价格
P0006C	Bradford蛋白浓度测定试剂盒(去垢剂兼容型)	800次	390.00元

碧云天生产的Bradford蛋白浓度测定试剂盒(去垢剂兼容型)(Detergent Compatible 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，能兼容一系列常见去垢剂，并且检测数据有很好的线性关系。本试剂盒的性能不仅优于常规的Bradford法，和BCA法比也更加快速便捷。
本试剂盒和常规的Bradford蛋白浓度测定试剂盒相比可以兼容一系列常见的去垢剂，和BCA法相比，能兼容高浓度的还原剂，并且检测速度极快，在检测含去垢剂样品蛋白浓度时，比BCA法更加便捷。生命科学研究中的很多蛋白样品都是含有去垢剂的，因此常规Bradford法的使用受到了很大的限制，通常只能使用可以兼容去垢剂但检测比较耗时的BCA法。如果使用本试剂盒，不仅能兼容去垢剂，而且由于能立即显色，检测速度会比BCA法快很多。当蛋白样品中同时含有一定浓度的去垢剂和还原剂时，常规的Bradford法和BCA法就都不能使用了，而本试剂盒仍然是可以检测的。
Bradford法的原理是考马斯亮兰(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。
本试剂盒测定蛋白浓度时不受较高浓度去垢剂的影响，可以很好地兼容1% Triton X-100、1% Tween 20、1% SDS、1%
NP-40和1% Brij35等去垢剂，同时兼容碧云天生产的多种裂解液，包括 Western及IP细胞裂解液(P0013)、RIPA裂解液(强)
(P0013B)、RIPA裂解液(中)(P0013C)、RIPA裂解液(弱)(P0013D)、NP-40裂解液(P0013F)、SDS裂解液(P0013G)、Western及
IP细胞裂解液(无抑制剂) (P0013J)、RIPA裂解液(强，无抑制剂) (P0013K) (参考图1)。
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，不仅能兼容上述的去垢剂，还能确保10μl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及其兼容性的实测效果图。A. 本试剂盒对于常见去垢剂的兼容性。B. 本试剂盒对于碧云天的多种裂解液的兼容性。蛋白标准品配制在图中所示的溶液中进行测试。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，本试剂盒测定蛋白浓度不受绝大部分样品中的化学物质的影响，尤其是还原试剂和去垢剂的影响。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。同时本试剂盒还能兼容上述各种常见的去垢剂。BCA法能耐受各种常见的去垢剂，但不能耐受高浓度的还原剂。
每个试剂盒可以检测800个样品。
包装清单：

产品编号	产品名称	包装
P0006C-1	G250染色液(去垢剂兼容型)	125ml×2
P0006C-2	蛋白标准(20mg/ml BSA)	1ml
—	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(去垢剂兼容型)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。为简便起见，如果蛋白样品所在溶液不含有去垢剂，也可以用0.9%NaCl、PBS或水稀释标准品。蛋白标准(20mg/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	92.5μl	20mg/ml BSA 7.5μl	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10μl不同浓度蛋白标准加到96孔板的蛋白标准孔中。
b. 取10μl样品到96孔板的样品孔中。如果样品不足10μl，需加标准品稀释液补足到10μl。请注意记录样品体积。
c. 各孔加入300μl G250染色液(去垢剂兼容型)。
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30分钟内测定，30分钟内检测数据无显著变化。对于不含去垢剂和含有某些去垢剂的情况，在2小时内检测数据无显著变化；对于含有某些特定去垢剂的情况，在2小时内检测数据会有一定的变化，但仍然会呈现较好的线性关系。
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：

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P0006	Bradford蛋白浓度测定试剂盒	1000次
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