An interesting paper in (Analytical Chemistry) describes the use of confocal fluorescence section imaging to analyze protein distribution in microarray spots on nitrocellulose film slides. By scanning and imaging through the Z dimension of a printed spot of fluor labeled protein, a 3-dimensional quantitation of fluorophore density in the films was obtained, showing that commercial … Continued
A SYSTEMS APPROACH TO QUANTITATIVE MULTIPLEXED RPPA’S Presented by: Michael A. Shultz, PhD RPPA Workshop in Kobe, Japan November, 2013 Porous nitrocellulose film (PNC) is the substrate of choice for RPPA largely because of its extraordinarily high protein binding capacity, which can exceed non-porous planar counterparts by 500X.
Results obtained from Western blots can be highly variable and dependent not only on the specific antigen/antibody pairs used, but also on other reagents in the protocol such as blocking buffer. Not surprisingly, no one “universal” blocking reagent works well with all proteins. Thus choice of blocking reagent should be assessed for optimal results and having … Continued