![]() Pierce ECL Western Blotting Substrate provides performance identical to the original Amersham ECL Western Blotting Detection Reagent from GE Healthcare. It enables the detection of picogram amounts of antigen and allows for easy detection of HRP using photographic or other imaging methods.īecause the luminol and peroxide reagent formulations are identical to other commercially available substrate products, one can switch to Pierce ECL without needing to optimize probing conditions or incubation protocols. Pierce ECL Western Blotting Substrate provides reliability and performance equivalent to other standard ECL substrates for detection of HRP enzyme activity. Recommended secondary antibody concentration: 1:1,000–1:15,000 dilution (0.07–1.0 µg/mL)Ĭompare and learn more about our western blot chemiluminescent substrates ›.Compatibility: nitrocellulose and PVDF membranes.Stability: 1-hr working solution stability, 1-year kit stability at 4☌.Pierce ECL Plus Western Blotting Substrate characteristics include: Expose blot to X -ray film or Chemiluminescence imaging system.Thermo Scientific Pierce ECL Western Blotting Substrate is a value-priced, entry-level horseradish peroxidase (HRP) substrate for enhanced chemiluminescence (ECL) that directly replaces costlier products without the need to re-optimize conditions. Cover blot with a clear plastic sheet protector or clear plastic wrap. HRP substrate is required per cm2 membrane area). Mix Detection Reagents 1 and 2 at a 1:1 ratio and add it to the blot (Approximately 0.1mL of working 5. Dilute the secondary antibody to 1:2000-1:20000 from a 1mg/mL stock. Dilute the primary antibody to 1:200-1:5000 from a 1mg/mL stock. Short-term exposure to typical laboratory lighting will not harm the working solution. For best results keep the substrate working solution in an amber bottle and avoid prolonged exposure to any intense light. Exposure to the sun or any other intense light can harm the substrate. Rust may cause speckling and high background. Metallic devices must have no visible signs of rust. ![]() All equipment must be clean and free of foreign material. Sodium azide is an inhibitor of HRP and could interfere with this system. Do not use sodium azide as a preservative for buffers. Add Tween™-20 Detergent (final concentration of 0.05-0.1%) to the blocking buffer and all diluted antibody solutions to minimize nonspecific signal.For optimal results, use a shaking platform during incubation steps. Using large blocking and wash buffer volumes minimizes nonspecific signal. Use sufficient volumes of wash buffer, blocking buffer, antibody solution and substrate working solution to cover blot and ensure that it never becomes dry. developed with Pierce ECL Western Blotting Substrate (Thermo Fisher. Avoid using milk as a blocking reagent when using avidin/biotin systems because milk contains variable amounts of endogenous biotin, which causes high background signal. Primary antibodies used in Western blot analysis were as follows: polyclonal anti. Blocking buffer also affects system sensitivity. Empirical testing is essential to determine the appropriate blocking reagent for each Western blot system, as cross-reactivity of the blocking reagent with antibodies causes nonspecific signal. To optimize antibody concentrations, perform a systematic dot blot analysis. Tanon High-sig ECL Western Blotting Substrate requires more dilute antibody concentrations than those used with precipitating colorimetric HRP substrates. To determine the appropriate concentrations, perform a systematic dot blot analysis. If you are currently using a SuperSignal Chemiluminescent Substrate, switching to Tanon High-sig ECL Western Blotting Substrate requires increasing antigen and antibody concentrations. Use the same blotting conditions when switching from Amersham ECL Substrate or Pierce ECL Substrate to Tanon High-sig ECL Western Blotting Substrate. The special formulation of Tanon High-sig ECL Western Blotting Substrate makes it the ideal substitute for Amersham ECL Substrate or Pierce ECL Western Blotting Substrate without the need for additional optimization of assay conditions. Blots can be repeatedly exposed to X-ray film or Chemiluminescence imaging system to obtain optimal results or stripped of the immunodetection reagents and re-probed. Tanon High-sig ECL Western Blotting Substrate enables the detection of picogram amounts of antigen and allows for easy detection of HRP using photographic or other imaging methods. Tanon™ High-sig ECL Western Blotting Substrate is a highly sensitive nonradioactive, enhanced luminol-based chemiluminescent substrate for the detection of horseradish peroxidase (HRP) on immunoblots. Tanon™ High-sig ECL Western Blotting Substrate Shaker / Oscillator / Mixer / Roller / Rotator.
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