Proteomics
Shotgun wedding: High-throughput elution of 2-D gels into multiwell plates
Fans of Billy Idol might recall that White Wedding, his 1982 smash hit, was about a "shotgun wedding" - an American colloquialism referring to a forced marriage between sometimes poorly matched partners. Proteomics has its own fair share of difficult relationships, one of these being between 2-D polyacrylamide gels and multiwell plates. A collaborative effort between the Agricultural Biotechnology Center in Hungary and the NIH has just resulted in a novel proteomic tool that is being termed "shotgun electro-elution" - an improved method for the direct, high-throughput electro-elution of 2-D gels without gel sectioning.
With the ever-increasing demand for massively parallel approaches in proteomics, the search is on for techniques that address existing challenges in sample throughput. One of the remaining bottlenecks in this regard is the transfer of protein spots from 2-D gels to multiwell plates for further analysis. Current high-throughput methods usually involve the slicing, piercing, or sectioning of gels by robotic workstations, followed by numerous sample extraction and elution steps prior to mass spectrometry. Although the direct elution of individual protein spots from intact 2-D gels has been possible for several years, several technical drawbacks still exist that have stymied the widespread use of this approach for high-throughput applications.
In an advance online publication in Electrophoresis, Zsuzsanna Buzás and colleagues describe an improved technique for the parallel (shotgun) elution of multiple protein spots into 384-well plates. An average of 70% recovery was achieved for several sample proteins as revealed by re-electrophoresis. Since this method has no moving components that contain liquids, it is uniquely suited for automation. One particularly interesting aspect of this approach is its apparent potential for use with 1536- and even 3456-well plates.
So, if you bring this particular shotgun into the proteomics lab, this is one marriage that might just last.
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