Pierce ECL Plus Western Blotting Substrate / ECL Plus蛋白质印迹底物
货号：32132,32132X3,32134
规格：100 mL,300 mL,25 mL
价格：3696,7614,1495
产品类型：免疫印迹/组化
品牌：Thermo Fisher
| Thermo Scientific的Pierce ECL Plus底物是一种用于印迹检测，基于9,10-二氢吖啶配方的化学发光及化学荧光HRP底物，可通过X射线胶片、CCD或激光成像仪进行成像。Pierce ECL Plus底物与HRP反应会生成吖啶酯，酯类中间体会与过氧化氢发生反应，产生强烈并持久的化学发光信号，这些信号可通过X射线胶片或CCD成像仪捕获。Pierce ECL Plus底物还可以产生强烈的荧光信号，通过荧光成像仪进行捕获。使用适当的一抗及二抗检测时，Pierce ECL Plus底物可检测硝化纤维素膜或PVDF膜上低皮克级的目标蛋白。Pierce ECL Plus底物具备动态范围宽、灵敏度高以及信号持续时间长等特点，是印迹分析应用的最佳选择。 |
| 特点： |
| - 简单易用— 可替代停产的GE ECL Plus底物，无需进行任何优化- 灵敏度更高— 可检测低皮克级的蛋白- 信号持续时间更长— 光信号持续时间长达5小时- 更多成像方法— 适用于X射线胶片、CCD或激光成像仪- 价格更经济— 相比其他品牌的类似产品，不仅具有高品质和高性能，同时价格也更低 |
| 数据： |
Obtain excellent detection of target proteins with ECL Plus SubstrateLysates from the cell lines indicated below were diluted in electrophoresis sample buffer and heated to 95°C for 5 minutes. Lane 1 contained 10μg of total protein, and seven 1:1 serial dilutions were loaded at 10μL/well. After electrophoresis, proteins were transferred to nitrocellulose membrane (Part No. 88013), and then the membranes were blocked with 5% milk in TBST. The membranes were incubated with primary antibody at 1:1000 dilution and then with the secondary antibody at 6.6ng/mL (1:150,000 dilution of 1mg/mL stock solution). A. Target: β-Catenin in HeLa cells; Primary Antibody: Rabbit Anti-β-Catenin (Thermo Scientific Lab Vision Part No. RB-1491-PABX); Secondary: HRP-conjugated Goat Anti-Rabbit IgG (Part No. 31460); B. Target: ERK1 in K562 cells; Primary Antibody: Mouse Anti-ERK1 (Part No. MA113041); Secondary Antibody: HRP-conjugated Goat Anti-Mouse IgG (Part No. 31430); C. Target: PP2A in A549 cells; Primary Antibody: Mouse Anti-PP2A (Millipore); Secondary Antibody: HRP-conjugated Goat Anti-Mouse IgG (Part No. 31430). CL-XPosure Film (Part No. 34090) and Pierce ECL Plus Substrate were used for detection. Exposure time was 5 minutes. Hsp86 was detected in HeLa cell lysates according to the following protocol. The lysates were diluted in electrophoresis sample buffer and heated to 95°C for 5 minutes. For each blot, lane 1 contained 10μg of total protein, and four 1:1 serial dilutions were applied to lanes 2-5 at 10μL/well. After electrophoresis, proteins were transferred to nitrocellulose membranes (Part No. 88013), and then the membranes were blocked with 5% milk in TBST. The membranes were incubated with Rabbit Anti-Hsp86 (Part No. PA3-013) at 1:1000 dilution and then with HRP-conjugated Goat Anti-Rabbit IgG (Part No. 31460) at 6.6ng/mL (1:150,000 dilution of 1mg/mL stock solution). (A) Blots were prepared using Pierce ECL Plus Substrate (Part No. 32132) or substrates from other suppliers, and then all blots were exposed to Thermo Scientific CL-XPosure Film (Part No. 34090). (B) Blots were prepared using Pierce ECL Plus Substrate and exposed to CL-XPosure Film at the indicated time points. (C) The membranes were prepared using Pierce ECL Plus Substrate and exposed to CL-XPosure Film for five seconds and scanned using the Syngene™ G:Box iChemiXT Imager (CCD; 1 minute exposure) and the Typhoon™ 9410 Variable Mode Imager (Laser-based imager; Excitation at 457nm, Emission at 510nm). |
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