WESTAR ECL SUBSTRATES FOR WESTERN BLOTTING

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WESTAR ECL Substrates for Western Blotting

Introduction

WESTAR is our product line of ECL HRP substrates for Western blotting.
Our double enhancer proprietary technology allows for modulation of signal intensity and signal duration.
Each WESTAR substrate is at the top of its respective market segment regarding performance/price ratio.

Features

All WESTAR substrates are:

  • Compatible with all chemiluminescence imagers and X-ray film detection
  • Well-suited for different types of membranes and blocking reagents as well as a wide range of antibody dilutions
  • Optimizados para garantizar un bajo background y una elevada relación señal-ruido

Chemiluminescence is the Western blotting detection method of election in most laboratories, as it provides the greatest sensitivity and convenience for detection with film or digital imaging equipment.
Chemiluminescent substrates for horseradish (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 components are mixed together.

In the chemiluminescent reaction, horseradish peroxidase catalyzes the oxidation of luminol into a reagent, which emits light when it decays. Since the oxidation of luminol is catalyzed by horseradish peroxidase, and the HRP is complexed with the protein of interest on the membrane, the amount and location of light that HRP catalyzes the emission of, is directly correlated with the location and amount of protein on the membrane.

However, the peroxide-catalyzed oxidation of luminol generates only a weak flash of light at 425 nm, which corresponds to visible blue light. The first generation of HRP detection substrates was based on the incorporation of a redox mediator or primary enhancer into the buffer, which was able to improve the enzyme turnover; thus increasing the equilibrium concentration of luminol radical anion, and thereby significantly improving the signal intensity. Recent publications have shown that by the addition of chemical compounds which act as secondary enhancers, the quality of the light emitted by the luminol oxidation can be furthermore increased, making it not only more intense but also steadier over time. Because of this breakthrough, the Italian company CYANAGEN S.r.l. has developed and patented a second generation of chemiluminescent substrates, called WESTAR.
All WESTAR substrates are protected by US7803573, EP1962095, US7855287, EP1950207, US2012009603 (A1), CA2742025, EP2405016, foreign equivalents and pending patents.

 

 

Product comparison.
Hela whole cell lysate (abcam®) was diluted to 250 µg/mL in electrophoresis reducing sample buffer and 1:1 serial dilutions were made. 5µL of each dilution (1250 ng to 78 ng of total protein) were separated by SDS-PAGE and the proteins transferred to Amersham™ Protran® Premium 0,45 NC membrane (GEHC). The membrane was blocked with 5% non-fat dry milk in 1X TBS TWEEN®20 Buffer. Then the membrane was incubated with Rabbit Anti-MAP Kinase (ERK 1/2) antibody (SIGMA) at 1:1000 followed by incubation with Anti-Rabbit IgG HRP Linked F(ab′)2 (GEHC) at 1:5000. WESTAR SUN was used for detection with Westar Reader (HI-TECH Cyanagen).

Product comparison.
Hela whole cell lysate (abcam®) was diluted to 250 µg/mL in electrophoresis reducing sample buffer and 1:1 serial dilutions were made. 5µL of each dilution (1250 ng to 78 ng of total protein) were separated by SDS-PAGE and the proteins transferred to Amersham™ Protran® Premium 0,45 NC membrane (GEHC). The membrane was blocked with 5% non-fat dry milk in 1X TBS TWEEN®20 Buffer. Then the membrane was incubated with Rabbit Anti-MAP Kinase (ERK 1/2) antibody (SIGMA) at 1:1500 followed by incubation with Anti-Rabbit IgG HRP Linked F(ab′)2 (GEHC) at 1:20000. WESTAR NOVA 2.0 was used for detection with Westar Reader (HI-TECH Cyanagen).

Product comparison.
Hela whole cell lysate (abcam®) was diluted to 250 µg/mL in electrophoresis reducing sample buffer and 1:1 serial dilutions were made. 5µL of each dilution (1250 ng to 78 ng of total protein) were separated by SDS-PAGE and the proteins transferred to Amersham™ Protran® Premium 0,45 NC membrane (GEHC). The membrane was blocked with 5% non-fat dry milk in 1X TBS TWEEN®20 Buffer. Then the membrane was incubated with Rabbit Anti-MAP Kinase (ERK 1/2) antibody (SIGMA) at 1:2000 followed by incubation with Anti-Rabbit IgG HRP Linked F(ab′)2 (GEHC) at 1:25000. WESTAR ETA C 2.0 was used for detection with Westar Reader (HI-TECH Cyanagen).

Product comparison.
Hela whole cell lysate (abcam®) was diluted to 250 µg/mL in electrophoresis reducing sample buffer and 1:1 serial dilutions were made. 5µL of each dilution (1250 ng to 78 ng of total protein) were separated by SDS-PAGE and the proteins transferred to Amersham™ Protran® Premium 0,45 NC membrane (GEHC). The membrane was blocked with 5% non-fat dry milk in 1X TBS TWEEN®20 Buffer. Then the membrane was incubated with Rabbit Anti-MAP Kinase (ERK 1/2) antibody (SIGMA) at 1:5000 followed by incubation with Anti-Rabbit IgG HRP Linked F(ab′)2 (GEHC) at 1:50000. WESTAR ETA C ULTRA 2.0 was used for detection with Westar Reader (HI-TECH Cyanagen).

Product comparison.
Hela whole cell lysate (abcam®) was diluted to 250 µg/mL in electrophoresis reducing sample buffer and 1:1 serial dilutions were made. 5µL of each dilution (1250 ng to 78 ng of total protein) were separated by SDS-PAGE and the proteins transferred to Amersham™ Protran® Premium 0,45 NC membrane (GEHC). The membrane was blocked with 5% non-fat dry milk in 1X TBS TWEEN®20 Buffer. Then the membrane was incubated with Rabbit Anti-MAP Kinase (ERK 1/2) antibody (SIGMA) at 1:10000 followed by incubation with Anti-Rabbit IgG HRP Linked F(ab′)2 (GEHC) at 1:100000. WESTAR SUPERNOVA was used for detection with Westar Reader (HI-TECH Cyanagen).

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