Amersham ECL™ Western Blotting Detection Reagents

Technical Information

Amersham™ ECL Western Blotting Detection Reagents were the world's first commercial chemiluminescent reagents for Western blotting and are still the most widely used. They are at least 10-fold more sensitive than colorimetric assays and produce excellent and reliable results every time.

These reagents use enhanced luminol-based detection which is suitable for all routine Western blotting. ECL Western Blotting Detection Reagents in combination with Hybond™ ECL membranes form a complete solution for your Western blot application.

Figure 1.
Target: rat brain homogenate Western blotted onto nitrocellulose (Hybond™ ECL, RPN 2020D); loadings: doubling dilutions starting at 1:20 dilution of the homogenate; detection: primary antibody - 1:5000 dilution of mouse anti-beta tubulin (N 357), secondary antibody - 1:5000 dilution of anti-mouse Ig HRP conjugate (NA 931) using ECL detection reagents (RPN 2106); exposure: on Hyperfilm™ ECL (RPN 2103) for 5 minutes.

The membrane is soaked briefly in the detection reagent. This elicits a peroxidase-catalyzed oxidation of luminol and subsequently enhanced chemiluminescence, where the HRP labeled protein is bound to the antigen on the membrane. The resulting light is detected on Hyperfilm ECL Western in minutes, or often seconds.

Contents include the following:

• RPN2109, 62.5 ml each of ECL Detection Reagents 1 and 2 (1000 cm² membrane)
  
• RPN2209, 125 ml each (2000 cm²)
  
• RPN2106, 250 ml each (4000 cm²)
  
• RPN2134, 375 ml each (6000 cm²)