SNAP i.d. Protein Detection System
Advantages of the SNAP i.d. System’s Vacuum Transport Feature
- Draws reagents through blotting membrane
- Minimizes overblocking
- Membranes are thoroughly flushed instead of just rinsed
- Reduced incubation times
Our proprietary blot holders and actively-driven reagent process ensure that the pores of the membrane are adequately blocked and washed.
Low concentrations of blocking reagent with SNAP i.d. improve quality.
Non-Fat Dry Milk (NFDM) is a blocking solution commonly used in Western blotting; however, its high blocking capacity may compromise the protein signal. To demonstrate this, a two-fold dilution series of rat liver lysate (12 µg in lane 1 to 0.09 µg in lane 8) was resolved with SDS-PAGE prior to blotting and immunodetection (the primary antibody was mouse anti-GAPDH; the secondary antibody was HRP conjugated goat anti-mouse). Blot A used a traditional immunodetection protocol (block for 1 hour in 5% NFDM, incubate in primary or secondary antibody at 1:50,000 for 1 hour, wash three times following incubations). Blots B and C were assembled in SNAP i.d blot holders and blocked for 20 s with either 0.5 or 0.05% NFDM, respectively. The blots were incubated for 10 m with anti-GAPDH (1:13,000), washed immediately and incubated for 10 m with HRP goat anti-mouse (1:10,000). Results show an increase in sensitivity with a decrease in milk concentration.
Improve your signal!Use low concentrations of blocking reagents with the SNAP i.d. system to improve quality.Non-Fat Dry Milk (NFDM) is an efficient blocking solution commonly used in western blotting; however, its high blocking capacity may compromise the protein signal. To demonstrate this, a two-fold dilution series of rat liver lysate (12 μg in lane 1 to 0.09 μg in lane 8) was resolved with SDS-PAGE prior to blotting and immunodetection. (The primary antibody was mouse anti-Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH); the secondary antibody was HRP-conjugated goat anti-mouse). Blot A used a standard immunodetection protocol (block for 1 hour in 5% NFDM, incubate in primary (1:40,000) or secondary antibody (1:50,000) for one hour, wash three times following incubations). Blot B, C and D were assembled in SNAP i.d blot holders and blocked for 20 seconds with either 0.5, 0.1 or 0.05% NFDM respectively. The blots were incubated for 10 minutes with anti-GAPDH (1:13,000), washed immediately and incubated for 10 minutes with HRP goat anti-mouse (1:10,000). Results show an increase in sensitivity with a decrease in milk concentration. |
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