Pierce G2 Fast Blotter

The Thermo Scientific Pierce G2 Fast Blotter is designed specifically for rapid semi-dry transfer of 10-300kDa proteins from polyacrylamide gels to nitrocellulose or PVDF membranes in 5 to 10 minutes.

The Pierce G2 Fast Blotter features an integrated power supply optimized to deliver consistent, high-efficiency protein transfer with commonly used pre-cast and homemade gels (SDS-PAGE) and nitrocellulose or PVDF membranes. The large cassette provides for simultaneous transfer of up to four mini-sized gels or two midi-sized gels. The blotter has an easy-to-use color LCD touchscreen interface and pre-programmed transfer methods for different numbers and sizes of gels and molecular weight ranges of proteins. The easy-touch programming feature allows custom transfer settings to be quickly created, saved and run.

The Pierce G2 Fast Blotter achieves highly efficient transfer in less than 10 minutes when used with Pierce 1-Step Transfer Buffer (Part No. 84731). The instrument is also effective for standard 30- to 60-minute semi-dry transfer protocols based on traditional buffers.

Highlights:

• Fast – transfer proteins in 5 to 10 minutes when used with 1-Step Transfer Buffer
• Efficient – achieve high transfer efficiency with a broad range of protein sizes (10-300kDa) compared to conventional semi-dry or wet (tank) transfer methods
• Integrated power supply – seamless operation between control unit and cassette provides consistent high efficiency protein transfer
• Easy-touch programming – access pre-programmed transfer methods or create, save and run customized transfer methods
• Convenient – simultaneously transfer 1 to 4 mini gels or 1 to 2 medium-sized gels
• Versatile – use with Pierce 1-Step Transfer Buffer for rapid blotting programs or Towbin transfer buffer for conventional semi-dry transfer methods

Includes:

• System consists of the Pierce G2 Fast Blotter Control Unit and Pierce G2 Fast Blotter Cassette; kit includes Western Blot Roller, Power Cord with C/13 Connector, and Quick Start Guide
• Unit ships with North American power cord; also available are country-specific power cords

Applications:

• Semi-dry Western blot transfer of 10-300kDa proteins from polyacrylamide gels to nitrocellulose or PVDF membranes

Product Details:

Traditional wet blotting techniques require transfer times of one hour to overnight to achieve good transfer efficiency. When used in conjunction with Pierce 1-Step Transfer Buffer, the Pierce G2 Fast Blotter is designed to provide transfer efficiency in 5-10 minutes that is equivalent, or better than, traditional transfer techniques for Western blotting. It also eliminates the need for gel pre-equilibration. This significant reduction in protein transfer time is accomplished by optimizing the ionic strength of the transfer buffer and increasing the current [amps (A)/cm2] flowing through the transfer stack. The Pierce G2 Fast Blotter can also be used for standard 30- to 60-minute semi-dry transfer protocols with Towbin buffer (Towbin et al., 1979).

Highly efficient transfer of low, medium, and high molecular weight proteins. The Pierce G2 Fast Blotter rapidly transfers a wide range of protein size from gel to membrane with similar or better efficiency compared to traditional transfer methods, such as tank and semi-dry, and other commercially available rapid blotting systems. HeLa lysate was serially diluted, prepared for SDS-PAGE, and electrophoresed according to the gel suppliers’ recommendations. Proteins were then transferred from gel to nitrocellulose or PVDF membrane using the specified methods and devices, following recommended protocols. Finally, blots were probed with respective specific antibodies, detected identically using Pierce Fast Western Blot Kit, SuperSignal West Dura Substrate (Part No. 35075) and imaged simultaneously using the Thermo Scientific myECL Imager (Part No. 62236).

Thermo Scientific Pierce G2 Fast Blotter provides highly efficient protein transfer in 10 minutes. GST-PI3K-SH2-HA (37kDa) was expressed in E. coli and purified with varying volumes of Thermo Scientific Anti-HA Magnetic Beads (Part No. 88836). Resulting samples were prepared for SDS-PAGE and electrophoresed. The proteins were then transferred from gel to nitrocellulose membrane using either traditional tank transfer (16 hours) with Towbin transfer buffer (25mM Tris, 139mM glycine, 20% methanol) or the Pierce G2 Fast Blotter (10 minutes) with Pierce 1-Step Transfer Buffer (Part No. 84731). Membranes were probed with anti-HA antibody for 1 hour, washed 5 times, probed with goat anti-mouse HRP for 30 minutes, washed 5 times and incubated in Thermo Scientific SuperSignal West Femto Substrate (Part No. 34096). The resulting blots were simultaneously imaged using the myECL Imager (Part No. 62236), and then band densitometry determined using myImageAnalysis Software (Part No. 62237).

References:

1. Towbin H. et al. (1979). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. USA. 76(9): 4350–54.

Related Resources:

Western Blot Tutorial Videos
Review of Western blot transfer methods

Related Products:

Country-specific and Replacement Power Cords
Pierce 1-Step Transfer Buffer
Western Blot Accessories
SuperSignal Western Blot Enhancer
myECL Imager
myImageAnalysis Software