WesternEaze-Chemi Kit

The WesternEaze-Chemi Kit includes all critical reagents required to generate high sensitivity Western blot data for 10 mini blots in one hour. The blocking buffer is specially formulated to increase sensitivity while decreasing overall background. The unique HRP Conjugate amplifies signal to increase sensitivity while minimizing the duration of incubation. The wash solution contains a gentle, non-ionic detergent that rapidly removes non-specifically bound material from the membrane without disrupting the antibody-antigen complex. WesternBright™ ECL substrate is included for sensitive, long-lasting and flexible detection using X-ray film or CCD camera. Provided as ready-to-use solutions designed to simplify your Western blot workflow.

Maximum sensitivity

Figure 1. High sensitivity Western blot in one-hour. Dilutions of HeLa lysate were electrophoresed using SDS-PAGE and the protein was transferred to a nitrocellulose membrane. The blots were probed with 5μg of mouse anti-Vinculin (Boster #MA1103) using either the WesternEaze or Traditional Western blot methods. The blots were developed with WesternBright™ ECL Substrate (Advansta #K-12045).

Optimized protocol for enhanced sensitivity

Figure 2. Alternate WesternEaze protocol significantly enhances sensitivity. Dilutions of Jurkat lysate were electrophoresed using SDS-PAGE and the proteins were transferred to nitrocellulose membranes. The detection sensitivity for rabbit monoclonal α-c-Myb (Abcam #ab109127) was determined using traditional, WesternEaze and the alternate WesternEaze blot methods. The blots were developed with WesternBright™ ECL Substrate (Advansta #K-12045). The alternate WesternEaze method enhanced sensitivity 36X compared to the standard WesternEaze method.

Reproducible results

Figure 3. WesternEaze data is highly reproducible. Dilutions of IFNα treated HeLa lysate were electrophoresed using SDS-PAGE and the proteins were transferred to nitrocellulose membranes. The three independent blots were each probed with 5μg of rabbit anti-phospho STAT3 (CST #4113S). The blots were developed with WesternBright™ ECL Substrate (Advansta #K-12045).

Comparable to traditional Western blot

Figure 4. Comparable results were generated with WesternEaze and traditional Western blot methods. Sample dilutions were electrophoresed using SDS-PAGE and the proteins were transferred to nitrocellulose membranes. The detection sensitivity for each target was determined using both a traditional method and the WesternEaze Method. The blots were developed with WesternBright™ ECL Substrate (Advansta #K-12045).

• WesternEaze-Chemi Kit brochure
• WesternEaze-Chemi Kit short protocol
• Buffer A, Eaze Blocking/Incubation Buffer SDS
• Buffer B, Eaze HRP Conjugate (Mouse or Rabbit) SDS
• Buffer C, Eaze Wash Buffer SDS
• WesternBright ECL SDS
• WesternBright Peroxide SDS
• Advansta’s Step-by-Step Guide to Western Blots