Pierce ECL Western Blotting Substrate

Thermo Scientific Pierce ECL Western Blotting Substrate is a value-priced, entry-level peroxidase substrate for enhanced chemiluminescence (ECL) that directly replaces costlier products without the need to re-optimize conditions.

Pierce ECL Western Blotting Substrate provides reliability and performance equivalent to other standard enhanced chemilumescent (ECL) substrates for detection of horseradish peroxidase (HRP) enzyme activity. Yet, Pierce ECL Substrate costs much less. Because 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. Simply switch to Pierce ECL and save a bundle.

Highlights:

• ECL substrate– an enhanced chemiluminescent substrate for detection of horseradish peroxidase (HRP) activity from antibodies and other Western blot probes
• About 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 at a significantly lower price than other companies (these claims are based on the 2010 U.S. list prices)
• No optimization required– switching to Pierce ECL substrate from other entry-level ECL substrates does not require optimization or protocol changes
• A name you can can rely on– count on the strong quality assurance, technical support and reputation behind Thermo Scientific Pierce Protein Research Products

Product Details:

Pierce ECL Western Blotting Substrate provides performance identical to the original Amersham ECL Western Blotting Detection Reagent from GE Healthcare, but it costs much less.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 are identical, they perform. the same.

Signal intensity of Thermo Scientific Pierce ECL Western Blotting 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.

Thermo Scientific Pierce ECL Western Blotting Substrate for detection of proteins on nitrocellulose or PVDF.β-actin and β-galactosidase protein in HeLa cell andE. colilysates were detected by Western blotting. The membranes were blocked with 5% skim milk and probed with primary antibody at 1µg/mL. The membranes were washed and then incubated with 0.2µg/mL of HRP-conjugated goat anti-mouse IgG (Part No. 31430) and then washed again. Working solutions of the Pierce ECL Substrate 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 and exposed to Thermo Scientific CL-XPosure Film (Part No. 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 (Part No. 88025). The membranes were blocked with 5% skim milk and probed with 4µg/mL of rabbit anti-bovine TNF-a(Thermo Scientific). The membranes were washed and then incubated with 0.4µg/mL of HRP-conjugated goat anti-rabbit IgG (Part No. 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 Resources:

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

Related Products:

PierceFastWestern Blot Kits with SuperSignal Substrates
Selection Guide for Chemiluminescent Western Blot Substrates
Blotting Equipment, Membranes and Signal Enhancers
CL-XPosure Film (X-ray film for chemiluminescence)