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ECL Western Blot Reagent

Introduction

Chemiluminescence is the western blot detection method of choice in most protein laboratories, as it provides the greatest sensitivity and convenience for detection with film or digital imaging equipment.

Chemiluminescent substrates for horseradish peroxidase (HRP) are two-component systems consisting of a stable peroxide solution and an enhanced luminol solution. To make a working solution, the equal volumes of the two components are mixed together. When incubated with a blot on which HRP-conjugated antibodies (or other probes) are bound, a chemical reaction produces light that can be detected by film or sensitive camera (Figure 1).

ABP Biosciences offers two types of super sensitive chemiluminescent substrates for Western blot detection with horseradish peroxidase enzyme (HRP). The signal from ECL substrates can be captured and documented using X-ray film or a capable CCD camera imager.

 
Figure1. The principle of WB detection.

ECL Western Blot Reagent Selection Guide

  Immobilon™ Western (Millipore) Super ECL Super ECL Ultra
 
Sensitivity picogram low picogram to high femtogram femtogram
Signal duration <2 h <8 h <4 h
Primary Ab dilution 1:1,000-1:20,000 1:1,000-1:30,000 1:5,000-1:30,000
Secondary Ab dilution 1:20,000-1:200,000 1:50,000-1:200,000 1:100,000-1:300,000
Stability 1 year at 4℃ 1 year at 4℃ 1 year at 4℃


Green_RedView Figure 1
The comparsion of sensitivity of Super ECL, Super ECL Ultra, and Millipore Immobilon™ Western blot substrate.


 
The comparsion on exposure time.


Green_RedView Figure 1 
The comparsion on signal duration.



The comparsion on antibody dilution.

To Order

Buy Cat. No. Product Name Description Unit Size Price
FP300 Super ECL Western Blotting Detection Kit ECL substrate 100 mL $120
FP301 Super ECL Western Blotting Detection Kit ECL substrate 500 mL $350
FP302 Super ECL Ultra Western Blotting Detection Kit Ultra sensitive ECL substrate  100 mL $200
FP303 Super ECL Ultra Western Blotting Detection Kit Ultra sensitive ECL substrate  200 mL $350