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Western检测

Quickblock™封闭液(PBS)
产品编号: P0220
产品包装: 100ml
产品价格: 116.00元
产品简介
使用说明
产品图片
相关产品
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产品问答
产品编号 产品名称 产品包装 产品价格
P0220 Quickblock™封闭液(PBS) 100ml 116.00元

      碧云天生产的QuickBlock™封闭液(QuickBlock™ Blocking Buffer)是最新一代的快速高效封闭液,总体效果显著优于传统的基于BSA(牛血清白蛋白)、脱脂奶粉、酪蛋白(Casein)等的封闭液及国外同类产品,可以用于Western blot (WB)、Immunofluorescence (IF)、Immunohistochemistry (IHC)、Immunocytochemitry (IC) 等实验中的封闭、一抗或二抗的稀释。本产品配制在PBS中,不含去垢剂。
      QuickBlock™封闭液快速高效。封闭时间通常仅需5-15分钟,并且和BSA、脱脂奶粉、酪蛋白等传统封闭液以及国外同类的快速 封闭液相比,显示出更强的信噪比(参考图1)。
      QuickBlock™封闭液封闭后背景极低。本封闭液不含血清和白蛋白,确保极高的信噪比。
      QuickBlock™封闭液兼容性好,兼容辣根过氧化物酶(Horseradish peroxidase, HRP)、碱性磷酸酶(Alkaline
phosphatase, AP)和生物素标记的二抗。本产品中添加了不影响HRP和AP活性的防腐剂,不会干扰HRP或AP标记二抗的检测。同时本产品不含生物素,不会干扰基于生物素的检测。
      QuickBlock™封闭液使用灵活。本产品配制在PBS中,可以直接用于相关实验,但也可以根据实验需要自行添加Tween-20或Triton X-100 至终浓度为0.05%-0.1%后用于封闭、一抗或二抗的稀释。
      本产品与BSA及国外同类产品的封闭效果对比参见图1。在相同样品和实验条件下,仅封闭液及封闭时间存在如下图所示的差异时,碧云天的 QuickBlock™封闭液封闭后的整体背景明显低于BSA封闭后的背景,而且目的条带亮度明显高于国外同类品牌产品。

      图1. QuickBlock™封闭液与BSA及国外同类产品的封闭效果比较。每组实验从左到右依次为:5μl蛋白Marker,2.5μg蛋白量的HeLa细胞裂解液,5μg蛋白量的HeLa细胞裂解液。请注意对于Antibody A,QuickBlock™封闭液比BSA和国外同类产品具有更低 的背景,并且和国外同类产品相比信号明显更强。对于Antibody B,QuickBlock™封闭液比BSA具有更低的背景,和国外同类产品相比信号明显更强。实际实验结果会因样品、抗体、实验条件等的不同而存在差异,图中数据仅供参考。


      关于不同封闭液的比较和选择,请参考碧云天的相关网页:http://www.beyotime.com/blocking- buffer.htm
      按照每张膜封闭需要5-10ml QuickBlock™封闭液计算,一个包装的本产品可以封闭10-20张膜。
包装清单:

产品编号 产品名称 包装
P0220 QuickBlock™封闭液(PBS) 100ml
说明书 1份

保存条件:
      4℃保存,一年有效。长期不使用可以-20℃保存。
注意事项:
      本产品推荐仅使用一次,重复使用可能会导致封闭效果下降。但对于一些信噪比很高的一抗,例如一些内参抗体,本封闭液可以重复使用2-3次。回收的封闭液请勿与未使用 过的封闭液混合。
      通常本产品用于PVDF膜及NC膜时的封闭时间为5-15分钟。对于一些背景非常高的抗体,可以尝试将封闭时间延长为30-60分钟。此外,如有特殊需要,也 完全可以4℃封闭过夜。
      由于没有任何一种封闭液是适用于所有实验体系的,因此对于一些特殊的实验,可能需要根据具体情况考虑使用其它更合适的封闭液。
      取放PVDF膜和NC膜应使用平头镊子,并仅轻轻夹取其边角,操作过程须避免膜表面产生划痕、折痕或压痕等痕迹。
      PVDF膜一经浸润和活化,需一直保持湿润,根据Western进行到的具体步骤可放置于western转膜液或洗涤液等适当溶液中,否则可能会产生难以封闭的异 常背景。
      在背景较高的情况下,为进一步提高信噪比,本产品中可以自行添加Tween-20或Triton X-100至终浓度为0.05%-0.1%
      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. Western Blot中膜的封闭
a. 完成转膜后,用Western洗涤液洗涤蛋白膜1-2分钟。
b. 根据膜的大小,在平皿或者其它适当容器中倒入一定体积的QuickBlock™封闭液(PBS)(可以根据实验需要自行添加Tween-20或Triton X-100至终浓度为0.05%-0.1%),确保封闭液后续能充分覆盖膜即可。对于常规的western,一张约6.6×8.5cm的膜推荐使用约10ml左右的封闭液。
c. 用平头镊夹住膜的一角,将膜放置在QuickBlock™封闭液(PBS)中,使封闭液完全浸没膜,置于水平摇床上封闭约10分钟(通常5-15分钟均可;经多种抗体的测试封闭10分钟的效果很多时候会显著优于常规的BSA封闭1小时的效果)
d. 封闭后的膜即可用于一抗孵育等后续实验。详细的Western操作可以参考如下的相关网页:
http://www.beyotime.com/western.htm
2. IF、IHC等实验的封闭
按照相关实验步骤,直接用QuickBlock™封闭液(PBS)替换传统封闭液即可(可以根据实验需要自行添加Tween-20或Triton X-100至终浓度为0.05%-0.1%),一般封闭时间可以缩短至10分钟(在碧云天测试过的多个一抗中,封闭10-20分钟的效果无明显差别,封闭10分钟的效果等同于或显著优于常规的封闭方法)。

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