Transfer proteins from gels to membranes in 10-20 minutes with conventional wet or semi-dry apparatus.  No additional equipment needed.

AMRESCO's Rapid Transfer Buffer offers fast, high efficiency, electrophoretic transfer of proteins from polyacrylamide gels to membranes for Western Blotting analyses.

Features:

• 10-20 minute transfer time versus traditional 1 hour to overnight transfer times.
• Transfers can be performed directly on the bench without cooling in contrast to conventional 1 hour to overnight transfer times that require cooling.
• Compatible with traditional wet and semi-dry transfer equipment - additional, expensive apparatus are not needed.
• Optimizes the sharpness of protein bands while reducing background and splotching on both PVDF and nitrocellulose membranes.
• Recommended for use with AMRESCO's RapidBlock™ for maximal signal to noise ratios on developed blots.
• The transfer buffer provided with AMRESCO's Rapid Western Blotting Kit.
• Methanol-free, non-hazardous formulation has no associated hazardous shipping charges and is an excellent substitute for traditional Towbin formulations for laboratories that want to reduce the use of hazardous reagents.

Semi-Dry Transfer
Cytoplasmic protein isolated from K562 cells using AMRESCO’s Cytoplasmic & Nuclear Protein Enrichment Kit (M330) was separated on a 10% Fluorescent Sprint Next Gel (M317) at 255 V for 25 minutes. Gels were washed with dH2O for 5 minutes and the image was captured using the Syngene GBox-HR Gel Doc System. Gels were equilibrated for 5 minutes in Towbin transfer buffer (1), Competitor 1 Transfer Buffer (2), or AMRESCO Rapid Transfer Buffer (3) and transferred with Bio-Rad’s Trans-Blot® SD Semi-Dry Electrophoretic Transfer Cell for at 300 mA for 50 minutes (1), 25 V for 10 minutes (2), or 25 V for 10 minutes (3). Image capture of the gels after transfer were performed with a Syngene GBox-HR Gel Doc System. The membranes were then probed with anti ß-actin antibody (1:1000) for 45 minutes and blots were developed with Visiglo™ HRP Plus Chemiluminescent Substrate Kit (N219).

Wet Transfer
10 µg of total protein isolated from K562 cells using Total Cell Protein Lysis Buffer (M329) was resolved on a 10% Fluorescent Sprint Next Gel (M317) at 250 V for 30 minutes. The gel image was captured using the Syngene GBox-HR Gel Doc System (1) prior to transfer to a PVDF membrane in a wet transfer apparatus at 90V/room temperature for 15 minutes with Rapid Transfer Buffer. After transfer the gel was stained with CoomassieR Blue, destained and documented with a Syngene GBox-HR Gel Doc System (2). The membrane was then probed with anti-β-tubulin antibody (1:5000) for 45 minutes with the Rapid Western Kit. The blot (3) was developed with Visiglo™ HRP Plus Chemiluminescent Substrate Kit (N219).

Specifications: