An entry-level ECL substrate at entry-level cost.

Thermo Scientific Pierce ECL Western Blotting Substrate performs identical to the Amersham ECL* Western Blotting Detection Reagent from GE Healthcare. The difference is that the Pierce ECL is half the cost of the leading competitor's product. Pierce ECL Western Blotting Substrate is an entry-level Western blotting substrate that is value-priced.

If you are currently using a needlessly expensive entry-level ECL substrate, you can switch to Pierce ECL Western Blotting Substrate without any optimization. Because these products perform the same, simply switch out the substrates and save a bundle.

• Half the cost of other ECL Substrates – low overhead and a commitment to customer value enables Thermo Fisher Scientific to make this product available to you for half of the cost other companies charge (these claims are based on the 2009 U.S. list prices)
• No optimization required – you can switch to our ECL substrate, from other entry-level ECL substrates, without the need for optimization or protocol changes
• A name you can can rely on – we put our strong technical support and reputation behind this product

Signal intensity of Thermo Scientific Pierce ECL Substrate is comparable to GE Healthcare ECL Substrate. HeLa cell lysate was separated by SDS-PAGE and transferred to nitrocellulose membrane to detect β-actin. The signal was detected and analyzed using the Kodak 1D Imaging Station. Error bars represent ±20% difference in relative intensity units.

Pierce ECL Substrate Western blot detection of proteins in lysate. Dilutions of HeLa cell E. coli lysate were prepared and separated by electrophoresis. The HeLa cell lysate proteins were transferred to nitrocellulose membranes (Product #88025) and bacterial lysate proteins were transferred to PVDF. The membranes were blocked with 5% skim milk and probed with primary antibody at 1 µg/ml (mouse anti-human Actin β, US Biological; mouse anti-β galactosidase AB-1, Lab Vision). The membranes were washed and then incubated with 0.2 µg/ml of HRP-conjugated goat anti-mouse IgG (Product #31430) and then washed again. Working solutions of the Pierce ECL and GE Healthcare ECL Reagent were prepared according to the manufacturers’ instructions and added to replicate membranes for 1 minute. The membranes were removed from the substrates and placed in plastic sheet protectors. Each membrane was exposed to CL-XPosure Film (Product #34090) and developed.

Pierce ECL Substrate Western blot detection of recombinant protein. Dilutions of recombinant bovine TNF-a (Thermo Scientific) were prepared, electrophoresed and transferred to nitrocellulose membranes (Product #88025). The membranes were blocked with 5% skim milk and probed with 4 µg/ml of rabbit anti-bovine TNF-a (Thermo Scientific). The membranes were washed and then incubated with 0.4 µg/ml of HRP-conjugated goat anti-rabbit IgG (Product #31460) and then washed again. Working solutions of the substrates were prepared according to the manufacturers’ instructions and added to the membranes for 1 minute. The membranes were removed from the substrates and placed in plastic sheet protectors. Each membrane was exposed to Hyperfilm Film (GE Healthcare).

Related Products
Chemiluminescent Western Blot Substrate Selection Guide – Compare Pierce ECL and SuperSignal substrates to determine which substrate or Western blotting kit is best for your application.
Membranes, Filter Paper and Transfer Buffer
CL-XPosure Film

Related Links
Tech Tip # 67: Chemiluminescent Western blotting technical guide and protocols
Tech Tip # 21: Convert to SuperSignal West Pico Substrate from ECL Substrate
Tech Tip # 22: Determine source of nonspecific background signal in Western blots
Tech Tip # 24: Optimize antigen and antibody concentrations for Western blots