Polyacrylamide gel photopatterning enables automated protein immunoblotting in a two-dimensional microdevice.
We demonstrate a two-dimensional microfluidic architecture that integrates polyacrylamide gel electrophoresis (PAGE) with immunoblotting in a fully automated format. This assay is designed to overcome performance limitations of conventional slab-gel immunoblotting, including multiple manual interventions, low-throughput transfer and blotting, and substantial consumption of reagents and sample. To unify PAGE with blotting in one device, this microfluidic approach makes use of high-resolution regional photopatterning of multiple polyacrylamide gel elements, and automated electrophoretic transport. A complete native immunoblot of free prostate specific antigen from human seminal fluid is demonstrated in less than 5 min. Further, the characterization of post-PAGE electrophoretic transfer showed high efficiency and minimal sample dispersion.
He
Mei
M
Department of Bioengineering, University of California, Berkeley, California 94720, USA.
Herr
Amy E
AE
D004591:Electrophoresis, Polyacrylamide Gel / Q000191:economics / Q000295:instrumentation*; D004867:Equipment Design; D006801:Humans; D015151:Immunoblotting / Q000191:economics / Q000295:instrumentation*; D008297:Male; D044085:Microfluidics / Q000191:economics / Q000295:instrumentation*; D017430:Prostate-Specific Antigen / Q000032:analysis* / Q000276:immunology; D012661:Semen / Q000737:chemistry*
D017430:Prostate-Specific Antigen