Save valuable time and material with RapidBlock™
RapidBlock™ Solution cuts an hour or more from Western blotting protocols by reducing blocking time to 5 minutes. Supplied as a 10X solution, RapidBlock™ is a protein-free formulation that provides very high signal to noise ratios with chemiluminescent substrates. Simply dilute and use.
Time course of blocking by RapidBlock™
Figure 1. Cytoplasmic proteins from K562 cells were resolved on a
12.5% Fluorescent Sprint ™ NEXT GEL™ (M318). Total protein and band resolution was determined by visualization on a UV transilluminator prior to transfer to
PVDF membrane. Membrane strips were blocked 5 or 30 minutes in either TBST/5% non-fat dry milk (lanes 1 and 2, respectively) or 1X RapidBlock™ Solution (lanes 3, 4 respectively). The strips were probed with anti-β-tubulin and detected with
VisiGlo™ HRP Plus (N219-Kit). Results demonstrate that 5 minute blocking in RapidBlock™ is sufficient to achieve a low background and an enhanced chemiluminescent signal compared to blocking in TBST/milk.
Chemiluminescent signal enhancement with RapidBlock™
Figure 2. 10 ug of cytoplasmic protein isolated from K562 cells with AMRESCO’s
Cytoplasmic/Nuclear Fractionation Kit (M330) was loaded per lane on a
12.5% Fluorescent Sprint Next™ Gel (M318) (Lanes A,B). Total protein and band resolution was determined by visualization on a UV transilluminator prior to transfer to PVDF membrane. The membrane was cut into strips and blocked 5 minutes in either TBST/5% non-fat dry milk (Lane 1) or AMRESCO’s RapidBlock™Solution (Lane 2). The blots were probed with β-tubulin antibody diluted in their respective blocking solutions, followed by washes with with TBS/0.1% Tween® 20 and incubation with HRP-conjugated secondary antibody diluted in their corresponding blocking solutions. Blots were washed prior to detection with
VisiGlo™ HRP Plus Chemiluminescent Substrate Kit (N219-Kit). Inset demonstrates the enhanced signal seen with 5 minutes of blocking in RapidBlock™.