Fluorescence detection is a preferred method for many quantitative protein microarray assays in research and diagnostic applications due to its excellent sensitivity and potential for multiplexing. Use of multiplexed fluorescence-based assays on porous nitrocellulose though has been hampered by high backgrounds at shorter excitation wavelengths (such as the 532 nm fluorescence channel).
To maximize the effectiveness of our premium ONCYTE Film Slides, we have developed a blocking reagent for improved performance of fluorescent endpoints. Super G Blocking Reagent allows for more robust fluorescence-based protein assays at multiple wavelengths such as the commonly used 532 nm and 635 nm channels.
Figure 1. Improved Fluorescence Background
Figure 1. Background fluorescence using Super G Blocking Reagent are 3- to 10-fold lower at 532 nm and up to 6-fold lower at 635 nm (Panel A). Typical blocking results with Super G are seen in Panel B where Super G blocking is clearly superior to PBST blocking.
Figure 2. Conversely, Signal-to-Noise is 4- to 10-fold higher at 532 nm (A) and up to 4.5-fold higher at 635 nm (B). Data presented are from SuperNOVA slides blocked with Super G or other protein- and non-protein-based blockers followed by sandwich assays for IL-1a, IL-1b, IL6, TNF b, and INFg. All data are shown relative to blocking with a common blocker (PBS with 0.1% Tween-20).