Thermo Scientific Pierce Western Blot Signal Enhancer is a two-reagent system for conditioning protein blots after transfer to greatly enhance the effectiveness of primary antibodies and intensify the final detection signal in Western blot experiments.
The Pierce Western Blot Signal Enhancer membrane treatment procedure is very simple, takes only 15 minutes and can be added to nearly any existing Western blotting protocol. The result is an increase in the intensity of target protein bands on the Western blot or detection of target proteins at levels that were previously not possible. The product is effective for signal intensification with both chemiluminescent and chromogenic substrates, especially with nitrocellulose membranes.
- Increases protein detection – most protein targets show a three- to 10-fold increase in signal intensity, enabling much less protein to be detected with the same substrate and method
- Improves antibody binding – the membrane-treatment reagent exposes and conditions target proteins so that specific antibodies can bind more effectively
- Works for nearly any protein – signal enhancement has been demonstrated with targets such as IL-6, p53, NFκB, BRCA1 and EGF
- Effective with any substrate – enhances both chemiluminescent (ECL) and colorimetric detection for Western blots
- Compatible with any membrane – enhances signal on nitrocellulose and PVDF membrane, regardless of pore size (enhancement is less pronounced with PVDF)
- Fast, 15-minute protocol – optimized for a combination of simplicity, speed and signal enhancement for most proteins
- Ready-to-use – no formulating or diluting necessary, and the reagents are stable for storage at room temperature
|Protocol summary for Thermo Scientific Pierce Western Blot Signal Enhancer.
||Enhanced chemiluminescent substrate detection using Thermo Scientific Pierce Western Blot Signal Enhancer. Identical gels and nitrocellulose blots of IL-6 protein samples (250 to 2000pg per well) were probed and detected with identical reagents and conditions after no treatment (top) or Enhancer-treatment (bottom) of the membrane. Final detection was with SuperSignal West Pico Chemiluminescent Substrate and exposure to film for 5 minutes.
||Enhanced chromogenic substrate detection using Thermo Scientific Pierce Western Blot Signal Enhancer. Identical gels and nitrocellulose blots of IL-6 protein samples (0.1 to 2µg per well) were probed and detected with identical reagents and conditions after no treatment (top) or Enhancer-treatment (bottom) of the membrane. Final detection was with NBT-BCIP substrate.
- Aupperlee, M., et al. (2005). Endocrinology 146, 3577-3588.
- Li, Y., et al. (2004). Blood. 104, 1137-1144.
- MacMillan, D., et al. (2005). J. Cell. Sci. 118, 5443-5451.
SuperSignal Western Blot Enhancer
SuperSignal West Pico Chemiluminescent Substrate
NBT-BCIP Substrate Solution