SignalFire? ECL Reagent

• 产品编号：CST-6883S      品牌：CST       原厂货号：6883S
• 产品分类：生化试剂 > 免疫分析试剂 > 免疫荧光溶液和试剂
• 应用分类：

描述：

该试剂是一种增强的发光氨系统，为配合CST™ HRP标记的二抗和Phototope ® -HRP 检测系统而设计。该试剂具有极高的敏感性，可以检测低达皮克级水平的蛋白。与其他的发光氨系统相比较， SignalFire™ ECL 试剂可以产生更强的信号，可以更长时间的输出信号。在存在过氧化物酶的情况下，辣根过氧化物酶（HRP）可以将发光氨转化成一种中度激活的二价阴离子。这种二价阴离子在返回其原状态时发光。底物暴露于HRP后，光发射立即达到最大并且可以持续0.5-1小时。光可以被记录在胶片上，一般暴露几秒钟。延长暴露时间可以增加灵敏度。

a）用TBS/T洗涤膜标记-HRP（结合抗体）3次，每次5分钟。 （b）用1体积的2X Reagent A和1体积的2X Reagent B配制成1X SignalFire™ ECL 试剂（如配置10ml，加5ml Reagent A 和5ml Reagent B ），混匀。 （c）将底物与膜孵育1分钟，去除多余的溶液（膜保持湿润），放入塑料盒中，用X片曝光。 避免皮肤反复暴露（请参考物品安全说明书或到我们的网站获取更多信息）

化学发光检测已经成为western blots检测中最好和最全面的检测方法。它们减少了放射性物质和有毒显色基质相关的危害。这些方法的检测速度和灵敏度的都是传统替代品无可比拟的。因为结果生成在胶片上，因此可以永久记录和保存实验结果，而且用化学发光方法检测出的印迹可以很容易被洗脱并且易于采用其它抗体重新检测。辣根过氧化物酶（HRP）结合的二抗与化学发光的底物特异性结合生成光信号。辣根过氧化物酶与抗体的轭合物有非常高的周转率，在较短的时间里有很高的敏感性。

原厂资料：

Description

SignalFire™ ECL Reagent from Cell Signaling Technology (CST) is an enhanced luminol based system designed for use with CST™ HRP-linked secondary antibodies and Phototope®-HRP detection assays. SignalFire™ ECL Reagent is extremely sensitive and capable of detecting protein levels at low picogram amounts. Compared to other luminol based systems, SignalFire™ ECL Reagent boasts a more robust signal and extended duration of signal output. In the presence of hydrogen peroxide, horseradish peroxidase (HRP) converts luminol to an excited intermediate dianion. This dianion emits light on return to its ground state. Light emission is maximal immediately after exposure of the substrate to HRP and continues for 1 hour. Light can be captured on X-ray film, typically by exposure for a few seconds. Maximum sensitivity can be obtained with longer exposure.

Directions for Use

(a) Wash membrane-bound HRP (antibody conjugate) three times for 5 minutes in TBS/T. 
 (b) Prepare 1X SignalFire™ ECL Reagent by diluting one part 2X Reagent A and one part 2X Reagent B (e.g. for 10 ml, add 5 ml Reagent A and 5 ml Reagent B). Mix well. 
 (c) Incubate substrate with membrane for 1 minute, remove excess solution (membrane remains wet), wrap in plastic and expose to X-ray film. 
 *Avoid repeated exposure to skin (see enclosed Material Safety Data Sheet or refer to our website for further information).

Background

Chemiluminescence systems have emerged as the best all-around method for western blot detection. They eliminate the hazards associated with radioactive materials and toxic chromogenic substrates. The speed and sensitivity of these methods are unequalled by traditional alternatives, and because results are generated on film, it is possible to record and store data permanently. Blots detected with chemiluminescent methods are easily stripped for subsequent reprobing with additional antibodies. HRP-conjugated secondary antibodies are utilized in conjunction with specific chemiluminescent substrates to generate the light signal. HRP conjugates have a very high turnover rate, yielding good sensitivity with short reaction times.

注意事项：

Storage: SignalFire™ ECL Reagent is stable at room tempera-
ture. DO NOT store Reagents A and B pre-mixed. Reagents A and
B should be combined just prior to exposing membranes