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Rapid Transfer Buffer

Transfer proteins from gels to membranes in 10-20 minutes with conventional wet or semi-dry apparatus

Rapid Transfer Buffer, 10X
Code: N789

10-20 minute transfer time versus traditional 1 hour to overnight transfer times
Transfer at room temperature
Methanol-free and non-hazardous
Compatible with traditional wet and semi-dry transfer equipment - additional, expensive apparatus are not needed

"We could successfully transfer 2D gels in 15 minutes..."

Samarjit Das, Ph.D.
Department of Cardiac Pathology
Johns Hopkins University
Baltimore, MD

Semi-Dry Transfer

Figure 1. 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

Figure 2. 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).

AMRESCO's Rapid Transfer Buffer offers fast, high efficiency, electrophoretic transfer of proteins from polyacrylamide gels to membranes for Western Blotting analyses. It is compatible with conventional wet and dry transfer apparatus and requires no additional equipment. Transfers with Rapid Transfer Buffer can be performed directly on the bench without cooling in contrast to conventional 1 hour to overnight transfer times that require cooling.

Rapid Transfer Buffer optimizes the sharpness of protein bands while reducing background and splotching on both PVDF and nitrocellulose membranes. It is recommended for use with AMRESCO's RapidBlock™ for maximal signal to noise ratios on developed blots. Rapid Transfer Buffer is the transfer buffer provided with AMRESCO's Rapid Western Blotting Kit.

The methanol-free, non-hazardous formulation of Rapid Transfer Buffer 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.