The Thermo Scientific Pierce 1-Step Transfer Buffer is designed for rapid semi-dry transfer of 10-300kDa proteins from polyacrylamide gels (SDS-PAGE) to nitrocellulose or PVDF membranes using the Pierce G2 Fast Blotter.
When used with the Pierce G2 Fast Blotter, this buffer provides effective gel-to-membrane transfer of proteins in 5 to 10 minutes with efficiency that is equal to, or better than, conventional Western blotting techniques. Pierce 1-Step Transfer Buffer is also compatible with other protein semi-dry transfer devices, including the original Pierce Fast Semi-Dry Blotter (Part No. 88217), when they are paired with a suitable high-current power supply. Such devices provide constant high current (1.3 to 5.0 amps) to rapidly transfer proteins via the high ionic strength conditions supplied by the transfer buffer.
- Fast – high ionic strength formulation provides 5- to 10-minute protein transfer when used with compatible fast semi-dry blotting systems
- Ready to use – supplied as a 1X solution that is stable and ready to use at room temperature
- Optimized – designed for seamless and reliable performance with the Pierce G2 Fast Blotter
- Compatible – effective with other semi-dry transfer devices equipped with a suitable high-current power supply, including the original Pierce Fast Semi-Dry Blotter (Part No. 88217)
- Economical – excellent performance without the special or costly consumables that are required by other fast semi-dry transfer devices
Pierce G2 Fast Blotter (Part No. 62288) or other semi-dry transfer device that is equipped or paired with a capable high-current power supply.
Power supply requirements for rapid semi-dry blotting using the Pierce 1-Step Transfer Buffer. For the given blot configuration and effective transfer area, the power supply must be able to deliver the indicated amps (A). This is based on 22mA per square centimeter of blot.
|1 mini gel
|2 mini gels or 1 midi gel
|3 mini gels
|4 mini gels or 2 midi gels
Fast blotting methods require a high-current power supply, such as the Pierce G2 Fast Blotter, and an optimized high ionic strength transfer buffer, such as Pierce 1-Step Transfer Buffer. By increasing the current, excellent transfer efficiency can be achieved in much shorter time compared to conventional methods. Amperage is held at a constant rate based on the surface area of the transfer stack(s) (~22-23mA/cm2) and voltage is limited to 25V.
|Rapid, high-efficiency semi-dry transfer with high ionic strength buffer. 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 (Part No. 62288); 10 minutes) with Pierce 1-Step Transfer Buffer. 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).
- Towbin H. et al. (1979). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences USA. 76(9): 4350–54.
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