Western Blotting, ECL

ACEsignal Chemiluminescent Substrate - EcoPlus

  • Catalog Number : A1040
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Introduction

ACEsignal Chemiluminescent Substrate – EcoPlus is a ready-to-use reagent for chemiluminescent detection of immobilized proteins (Western blotting), conjugated with horseradish peroxidase (HRP) directly or indirectly. In the presence of hydrogen peroxide, HRP catalyzes theoxidation of cyclic diacylhydrazides, such as luminol, and light emits. ACEsignal Chemiluminescent Substrate – EcoPlus provides a convenient way to visualize HRP-based detection. Simply mix and add the solutions onto the membrane. The signal of target proteins can be recorded by exposure to X-ray films or compatible image acquisition system.

General Information

Storage instruction ACEsignal Chemiluminescent Substrate – EcoPlus should be stored at 2-8 °C and shielded from light. Expiration date is labeled on

MATERIALS NEEDED BUT NOT PROVIDED

  1. PVDF or nitrocellulose membrane
  2. Wash buffer: Phosphate-buffered saline (PBS) or Tris-buffered saline (TBS) containing 0.05–0.1% Tween®-20
  1. PBS: 10 mM KH2PO4,150 mM NaCl, pH 7.4
  2. TBS: 25 mM Tris, 150 mM NaCl, pH 7.4
  1. Blocking buffer: 1–5% (w/v) blocking agent (e.g., casein, BSA, or gelatin) in wash buffer
  2. Specific primary antibody for interested protein, diluted in blocking buffer
  3. HRP-conjugated secondary antibody, specific for primary antibody, diluted in blocking buffer
  4. X-ray film or chemiluminescence image acquisition systems

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