ONE-HOUR WesternTM Basic Kit (Rabbit) (5 assays/kit)

ONE-HOUR Western^TM Basic Kit (Rabbit)

Kit Description

GenScript's ONE-HOUR Western^TM Kit yields journal-quality Western or Dot blot results in about one hour. Using GenScript's breakthrough immunodetection technology (patent pending), the kit replaces the classical three-step Western process, which take nearly five hours. Transfer the proteins from gel to membrane and incubate it in the pretreat solution for five minutes. Then incubate in WB-2 solution with the mixture of primary antibody and WB-1 solution for 40 minutes. Then wash three times for five or ten minutes each. The membrane can then be developed with the HRP substrate. The ONE-HOUR Western^TM procedure is contrasted with a classical three-step Western at right. 

The ONE-HOUR Western^TM Kit is designed to produce high signal with low background for quick and clear Western analysis of proteins. 

Except for basic kits, all other kits contain all the necessary reagents, buffers and materials such as membrane for performing a Western blot, and no additional secondary antibody is needed. 

The ONE-HOUR Western^TM Kit saves precious research time. For added cost-efficiency, we recommend our complete kits, which contain all the reagents, membrane, HRP substrate necessary for a perfect blot. However, the kit components are also sold separately. 

Key Features

• Easy to perform: This kit has fewer and simpler steps than other Western kits.
• Low background: The kit contains WestClear^TM Nitrocellulose Membrane and LumiSensor^TM series of Chemiluminescent HRP Substrate Kits optimized for low background.
• Reproducible results: The kit produces highly reproducible results.
• No additional secondary antibody is needed.

Examples:

Comparison of ONE-HOUR Western^TM blot detection with classical Western blot detection.
ONE-HOUR Western^TM blot detection was compared with classical Western blot detection of α-Tubulin protein from Hela cell lysate. Serially diluted Hela cell lysate samples were Western blotted to WestClear^TM Nitrocellulose Membrane (GenScript, L00224) after SDS-PAGE. The membrane was then cut into two halves and processed with different procedures using α-Tubulin monoclonal antibody (abcam, ab7291): ONE-HOUR Western^TM blot detection (1 hr, right panel of Figure 2) and classical Western-blot detection (4.5 hours, left panel of Figure 2).

Figure 2. Western blots for the detection of α-Tubulin protein from Hela cell lysate by both classical Western and ONE-HOUR Western^TMBasic Kit. 10 μg, 2.5 μg, 0.63 μg, and 0.16 μg of Hela cell lysate (BD Biosciences, 611449) were loaded in different lanes as shown in the figure. The classical Western blot was developed with ECL system (GE Healthcare, RPN2109). The ONE-HOUR Western blot was developed with LumiSensor^TM Chemiluminescent HRP Substrate Kit (GenScript, L00221V60).

Technical Manual

Components