描述：
CST公司的SignalFire™ Plus ECL Reagent是一种极敏感的化学发光底物，可通过western blot检测低皮克级别的蛋白质。SignalFire™ Plus 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™ Plus 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。储存：保存于4ºC，这个产品在12个月中稳定。请勿将Reagent A 和Reagent B 混合保存。 Reagent A 和Reagent B 在膜曝光前使用。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™ Plus ECL Reagent from Cell Signaling Technology (CST) is a highly sensitive chemiluminescent substrate capable of detecting low picogram amounts of protein by western blot analysis. SignalFire™ Plus ECL Reagent has an extended duration of signal output lasting several hours following blot exposure, allowing for multiple exposures with either film or a digital imaging system. The strong signal output allows detection of low abundance proteins, conservation of reagents, and short exposure times.
SignalFire™ Plus ECL Reagent requires approximately five-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.
CST公司的SignalFire™ Plus ECL Reagent是一种极敏感的化学发光底物，可通过western blot检测低皮克级别的蛋白质。SignalFire™ Plus 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™ Plus 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。
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™ Plus 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.
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