Although Super G blocking reagent was developed and optimized for fluorescent endpoint detection on nitrocellulose film slides, we have found that it is also an effective blocker for Western blot applications. Data presented below illustrate the effectiveness of Super G blocking reagent in typical Western blot experiments using nitrocellulose membranes by reducing background staining and allowing for clear detection of protein biomarkers.
Figure 1. Improved Fluorescence Backgrounds for Protein Arrays
||Figure 2. Excellent Background Levels for Colorimetric Endpoint Detection on Western Blots|
Figure 1. Typical results achieved with Super G blocking reagent compared to PBST blocking on a protein array using fluorescence detection. 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 on nitrocellulose film slides (ONCYTE SuperNOVA).
Figure 2. Typical results achieved with Super G blocking reagent compared to casein blocking on Western blots using colorimetric endpoint detection. Per panel, 625 ng (left lane) and 2500 ng (right lane) of mouse liver lysates were separated by SDS-PAGE and transferred to Whatman Protran nitrocellulose membranes. Membranes were blocked for 1 hour with respective blockers followed by detection with anti-GAPDH antibody (1:1000) with colorimetric endpoint detection utilizing alkaline phosphatase with BCIP/NBT substrate (VECTASTAIN ABC System).