Western blot blocking in 5 minutes

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.

• Chemiluminescent signal enhancement
• Block PVDF or nitrocellulose membranes in 5 minutes
• Enhance chemiluminescent signals
• Reduce background and cross-reactivity

Time course of blocking by RapidBlock™

Cytoplasmic proteins from K562 cells were resolved on a 12.5% Fluorescent Sprint ™ NEXT GEL™ (AMR-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 (AMR-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™

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 (AMR-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 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 (AMR-N219-Kit). Inset demonstrates the enhanced signal seen with 5 minutes of blocking in RapidBlock™.