SignalFire? Elite ECL Reagent

• 产品编号：CST-12757S      品牌：CST       原厂货号：12757S
• 产品分类：生化和细胞分析 > 标签、标记试剂盒 > 酶底物 > 化学发光底物
• 应用分类：

描述：

CST公司的SignalFire™ Elite ECL Reagent是一种极敏感的化学发光底物，可通过western blot检测飞克级别的蛋白质。SignalFire™ Elite ECL Reagent与胶片和数码照片兼容。极其强烈的信号输出允许检测低丰度蛋白,保护试剂,缩短曝光时间。SignalFire™ Elite ECL Reagent相较于传统ECL试剂只需要少于1/10的Anti-rabbit IgG, HRP-linked Antibody #7074 or Anti-mouse IgG, HRP-linked Antibody #7076。限制HRP膜的曝光量可以防止高背景、靶蛋白信号的过饱和,或假阴性结果。其他HRP偶联抗体,包括HRP偶联一抗和anti-biotin-HRP抗体,应同样被稀释。稀释二抗可能需要优化。溶菌产物和初级抗体浓度的滴定建议实现最佳信噪比。（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）化学反光检测系统已经成为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与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™ Elite ECL Reagent from Cell Signaling Technology (CST) is an ultra sensitive chemiluminescent substrate capable of detecting femtogram amounts of protein by western blot analysis. SignalFire™ Elite ECL Reagent is compatible with both film and digital imaging systems. The extremely intense signal output allows detection of very low abundance proteins, conservation of reagents, and short exposure times.

SignalFire™ Elite ECL Reagent requires approximately ten-fold less Anti-rabbit IgG, HRP-linked Antibody #7074 or Anti-mouse IgG, HRP-linked Antibody #7076 than traditional ECL reagents. Limiting the amount of HRP exposed to the membrane prevents high background, oversaturation of the target protein signal, or false negative results. Other HRP-conjugated antibodies, including HRP-conjugated primary and anti-biotin-HRP antibodies, should be diluted similarly. Dilution of secondary antibody from alternative vendors may need to be optimized. Titration of lysate and primary antibody concentration is recommended to achieve optimal signal-to-noise ratio.

Bioactivity

Storage: Store at 4ºC. This product is stable for 12 months. DO NOT store Reagents A and B pre-mixed. Reagents A and B should be combined just prior to exposing membranes.

Directions for Use

(a) Wash membrane-bound HRP (antibody conjugate) at least three times for 5 minutes in TBS/T. It is very important to thoroughly wash the membrane prior to substrate incubation.

(b) Prepare 1X SignalFire™ Elite 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: Store at 4ºC. This product is stable for 12 months. DO
NOT store Reagents A and B pre-mixed. Reagents A and B should
be combined just prior to exposing membranes