生物素标记EMSA探针-NF-κB (0.2μM)(GS056B)

生物素标记EMSA探针-NF-κB (0.2μM)

产品编号: GS056B

产品包装:200μl
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200μl

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675.00 10

价格: ¥ 675.00

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产品编号 产品名称 产品包装 产品价格
GS056B 生物素标记EMSA探针-NF-κB (0.2μM) 200μl 675.00元

      生物素标记EMSA探针-NF-κB是用于EMSA(也称gel shift)研究的并经生物素(Biotin)标记的NF-κB consensus
oligonucleotide。这个生物素标记的双链寡核苷酸含有公认的NF-κB结合位点,可以用作EMSA研究时的探针。
      NF-κB consensus oligo的序列如下:
      5′-AGT TGA GGG GAC TTT CCC AGG C-3′
      3′-TCA ACT CCC CTG AAA GGG TCC G-5′
      本生物素标记EMSA探针已经过纯化,可以直接用于EMSA结合反应。
      本生物素标记EMSA探针可以和碧云天的化学发光法EMSA试剂盒(GS009)配套使用。
      一个包装的生物素标记探针可以进行约200-400个样品的EMSA检测。
包装清单:

产品编号 产品名称 包装
GS056B 生物素标记EMSA探针-NF-κB(0.2μM) 200μl
说明书 1份

保存条件:
-20℃保存,一年有效。
注意事项:
避免加热到40℃以上,温度过高会导致双链DNA探针解聚成单链。而单链无法用于EMSA研究。
对于基于生物素标记的EMSA检测的详细操作可以参考碧云天的化学发光法EMSA试剂盒(GS009)的使用说明。
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. 本生物素标记EMSA探针用于EMSA结合反应时,参考如下步骤进行:
A. 如下设置EMSA结合反应:

阴性对照反应:
Nuclease-Free Water 7-7.5μl
EMSA/Gel-Shift 结合缓冲液(5X) 2μl
细胞核蛋白或纯化的转录因子 0μl
生物素标记探针 0.5-1μl
总体积 10μl
样品反应:
Nuclease-Free Water 5-5.5μl
EMSA/Gel-Shift 结合缓冲液(5X) 2μl
细胞核蛋白或纯化的转录因子 2μl
生物素标记探针 0.5-1μl
总体积 10μl
探针冷竞争反应:
Nuclease-Free Water 4-4.5μl
EMSA/Gel-Shift 结合缓冲液(5X) 2μl
细胞核蛋白或纯化的转录因子 2μl
未标记的探针 1μl
生物素标记探针 0.5-1μl
总体积 10μl
突变探针的冷竞争反应:
Nuclease-Free Water 4-4.5μl
EMSA/Gel-Shift 结合缓冲液(5X) 2μl
细胞核蛋白或纯化的转录因子 2μl
未标记的突变探针 1μl
生物素标记探针 0.5-1μl
总体积 10μl
Super-shift反应:
Nuclease-Free Water 4-4.5μl
EMSA/Gel-Shift 结合缓冲液(5X) 2μl
细胞核蛋白或纯化的转录因子 2μl
目的蛋白特异抗体 1μl
生物素标记探针 0.5-1μl
总体积 10μl

注:生物素标记EMSA探针的推荐用量为每个反应0.5微升,如果检测出来的目的蛋白的EMSA条带偏弱,可以适当加大生物素标记EMSA探针的用量至0.75微升或1微升。
注2:对于冷竞争时使用的未标记的探针或未标记的突变探针,使用量可以根据实际情况调整使用的体积。推荐的用于冷竞争的未标记的探针或突变探针的用量为生物素标记探针的50-100倍。
B.按照上述顺序依次加入各种试剂,在加入标记好的探针前先混匀,并且室温(20-25℃)放置10分钟,从而消除可能发生的探针和蛋白的非特异性结合,或者让冷探针优先反应。然后加入标记好的探针,混匀,室温(20-25℃)放置20分钟。
C.加入1μl EMSA/Gel-Shift上样缓冲液(无色,10X),混匀后立即上样。注意:有些时候溴酚蓝会影响蛋白和DNA的结合,建议尽量使用无色的EMSA/Gel-Shift上样缓冲液。如果对于使用无色上样缓冲液在上样时感觉到无法上样,可以在无色上样缓冲液里面添加极少量的蓝色的上样缓冲液,至可以观察到蓝颜色即可。
2.对于基于生物素标记的EMSA检测的更多详细操作可以参考碧云天的化学发光法EMSA试剂盒(GS009)的使用说明。

生物素标记EMSA探针-NF-κB (0.2μM)(GS056B)

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